Analogs of fexaramine and methods of making and using

ABSTRACT

Novel compounds having a formula 
     
       
         
         
             
             
         
       
     
     embodiments of a method of making the same, and of a composition comprising them are disclosed herein. Also disclosed are embodiments of a method of treating or preventing a metabolic disorder in a subject, comprising administering to a subject (e.g., via the gastrointestinal tract) a therapeutically effective amount of one or more of the disclosed compounds, thereby activating FXR receptors in the intestines, and treating or preventing a metabolic disorder in the subject. Additionally disclosed are embodiments of a method of treating or preventing inflammation in an intestinal region of a subject, comprising administering to the subject (e.g., via the gastrointestinal tract) a therapeutically effective amount of one or more of the disclosed compounds, thereby activating FXR receptors in the intestines, and thereby treating or preventing inflammation in the intestinal region of the subject.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation-in-part of International ApplicationNo. PCT/US2015/020552, filed Mar. 13, 2015, which claims the benefit ofU.S. Provisional Application No. 61/952,763 filed Mar. 13, 2014 and62/061,607 filed Oct. 8, 2014. This application also claims the benefitof U.S. Provisional Application No. 62/252,045, filed Nov. 6, 2015. Eachof these prior applications is incorporated herein by reference.

ACKNOWLEDGMENT OF GOVERNMENT SUPPORT

This invention was made with government support under Grant No.R24-DK090962 awarded by the National Institute of Health (NIH). Thegovernment has certain rights in the invention.

FIELD

This disclosure concerns new fexaramine analogs and a method for usingthe analogs to treat or prevent gastrointestinal (GI) inflammatoryconditions, intestinal permeability conditions, intestinal alteredmicrobiome conditions, cholestatic disorders, bile disorders, intestinalabsorption disorders, and metabolic disorders, including obesity anddiabetes.

PARTIES TO JOINT RESEARCH AGREEMENT

Salk Institute for Biological Studies and The University of Sydney areparties to a joint research agreement governing inventions disclosedherein.

BACKGROUND

Metabolic syndrome, a western diet-induced, pro-inflammatory diseaseaffecting up to 25% of Americans, is characterized by central obesity,impaired glucose tolerance, dyslipidemia, insulin resistance, and typeII diabetes. Secondary complications associated with metabolic syndromeinclude atherosclerosis, stroke, fatty liver disease, blindness,gallbladder disease, cancer, polycystic ovary disease and others.Consequently there is interest in reducing food intake, losing weight,and reducing elevated blood glucose. There is also an interest incombating obesity and related conditions using methods that do notrequire drastic lifestyle or dietary changes. In addition, inflammatorygastrointestinal conditions resulting from various types of pathologyaffect millions of people. Thus, effective and targeted treatments forvarious inflammatory gastrointestinal (GI) conditions are also needed.

Farnesoid X receptor (FXR) is a ligand-activated transcriptionalreceptor expressed in diverse tissues including the adrenal gland,kidney, stomach, duodenum, jejunum, ileum, colon, gall bladder, liver,macrophages, and white and brown adipose tissue (Forman et al., Cell81:687-693 (1995). FXR has been reported to contribute to the regulationof whole body metabolism including bile acid/cholesterol, glucose andlipid metabolism. Synthetic ligands for FXR have been identified andapplied to animal models of metabolic disorders, but these knownsynthetic ligands have shown limited efficacy and, in certain cases,exacerbated phenotypes.

Bile acids (BAs) function as endogenous ligands for FXR such thatenteric and systemic release of BAs induces FXR-directed changes in geneexpression networks (Lee et al., Trends Biochem Sci 31:572-580, 2006;Repa et al., Science 289:1524-1529, 2000; Zollner et al., J Hepatol39:480-488, 2003; Fang et al.,. J Biol Chem 283:35086-35095, 2008;Kemper et al., Cell Metab 10:392-404, 2009; Makishima et al., Science284:1362-1365, 1999; Stedman et al., Proc Natl Acad Sci U S A103:11323-11328, 2006). The complex role of FXR in metabolic homeostasisis evident in studies on whole body FXR knockout (FXR KO) mice. On anormal chow diet, FXR KO mice develop metabolic defects includinghyperglycemia and hypercholesterolemia, but conversely, exhibit improvedglucose homeostasis compared to control mice when challenged with a highfat diet (Sinal et al., Cell 102:731-744, 2000; Prawitt et al., Diabetes60:1861-1871, 2011). Similar contrary effects are seen with systemic FXRagonists, with beneficial effects observed when administered to chow-fedmice and exacerbated weight gain and glucose intolerance observed whenadministered to diet-induced obesity (DIO) mice (Zhang et al., Proc NatlAcad Sci U S A 103:1006-1011, 2006; Watanabe et al., J Biol Chem286:26913-26920, 2011). In the liver, FXR activation suppresses hepaticBA synthesis, alters BA composition, reduces the BA pool size (Wang etal., Dev Cell 2:721-731, 2002; Fang et al., Mol Cell Biol 27:1407-1424,2007; Lu et al., Mol Cell 6:507-515, 2000), and contributes to liverregeneration (Huang et al., Science 312:233-236, 2006) as well as lipidand cholesterol homeostasis (Zhang et al., Genes Dev 18:157-169, 2004;Ma et al., J Clin Invest 116:1102-1109, 2006). Consistent with this,activation of hepatic FXR by the synthetic bile acid 6α-ethylchenodeoxycholic acid (6-eCDCA) is beneficial in the treatment ofdiabetes, non-alcoholic fatty liver disease (NAFLD), and primary biliarycirrhosis (PBC) (Stanimirov et al., Acta Gastroenterol Belg 75:389-398,2012; Mudaliar et al., Gastroenterology 145:574-582 e571, 2013).

FXR is also widely expressed in the intestine where it regulatesproduction of the endocrine hormone FGF15 (FGF19 in humans), which, inconjunction with hepatic FXR, is thought to control BA synthesis,transport and metabolism (Kim et al., J Lipid Res 48:2664-2672, 2007;Song et al., Hepatology 49,:97-305, 2009; Inagak et al., Cell Metab2:217-225, 2005). Intestinal FXR activity is also known to be involvedin reducing overgrowth of the microbiome during feeding (Li et al., NatCommun 4:2384, 2013; Inagaki et al., Proc Natl Acad Sci U S A103:3920-3925, 2006).

SUMMARY

In view of the above, there is an ongoing need for methods andcompositions for the treatment and prevention of metabolic disorders,including obesity and metabolic syndrome. As well as other disorderssuch as gastrointestinal (GI) inflammatory conditions, intestinalpermeability conditions, intestinal altered microbiome conditions,cholestatic disorders, intestinal absorption disorders, and biledisorders. There is also a need for methods and compositions thatproduce beneficial clinical effects, while reducing side effects, suchas those resulting from systemic administration of a particular therapy(such as systemic FXR-directed therapies). There also is a need forcompositions that specifically target intestinal FXR, which can resultin a beneficial anti-inflammatory effect in the intestines. Disclosedembodiments of the present disclosure address these needs, and providenovel compounds and compositions that target intestinal FXR. In someexamples, the compounds are gut-selective, non-bile acid FXR agonists.

Certain disclosed compounds have the following general formula

With reference to the general formula, R is selected from

R^(a) is selected from aryl, heteroaryl, alkyl, alkenyl, cycloalkyl,heterocyclic, or polycyclic; R^(b) is selected from hydrogen, alkyl,alkenyl, or cycloalkyl; Y is CR^(g), N or N—O (N-oxide); R^(c), R^(d),R^(e) and R^(g) are each independently selected from hydrogen,deuterium, halide, alkyl, alkenyl, alkoxy, alkylthio, amino, sulfonyl,aminosulfonyl, aminocarbonyl, acyl, hydroxyl or nitro; R^(fa) and R^(fb)are each independently selected from hydrogen, deuterium, halide oralkyl; L^(a) and L^(b) are each independently selected from hydrogen,deuterium, alkyl or cycloalkyl, or together form a pi-bond; L^(c) andL^(d) are each independently selected from hydrogen, deuterium, alkyl orcycloalkyl; W is selected from O or —(C(L^(c))(L^(d)))_(s)-; s is 1, 2,3, 4, 5 or 6;n is 0 or 1; and X is aryl, heterocyclic or heteroaryl.

Also with reference to the general formula the following provisos apply:

-   if W is CH₂ and L^(c) and L^(d) are both H, then X is not a    benzopyran;-   if R is

L^(c) and L^(d) are both H, and L^(a) and L^(b) are both H or togetherform a pi-bond, then X is not a benzopyran;

-   X is not substituted with —R^(x)-L^(x)-R^(x2), where

R^(x) is selected from O, NR^(x3), sulfonyl or S;

R^(x3) is selected from H, alkyl, alkenyl, alkynyl, cycloalkyl,cycloalkenyl, or aryl;

L^(x) is selected from a bond, alkylene, alkenylene, alkynylene,cycloalkyl, cycloalkenyl, heterocyclic, aryl, heteroaryl orCR^(x4)R^(x5);

R^(x4) and R^(x5) are each independently selected from H, D, halogen,alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, —C(O)OR^(x6), or—C(O)NR^(x6)R^(x7);

R^(x6) and R^(x7) are each independently selected from H, alkyl,alkenyl, alkynyl, cycloalkyl or cycloalkenyl;

R^(x2) is selected from —C(O)L^(x2)R^(x8) or a carboxyl bioisostere;

L^(x2) is a bond or NR^(x3);

R^(x8) is H, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl,—OR^(x9), N(R^(x9))₂, —C(O)R^(x9), —S(O)₂R^(x9), —C(O)OR^(x9),—S(O)₂N(R^(x9))₂ or C(O)N(R^(x9))₂; and

each R^(x9) is independently selected from H, alkyl, alkenyl, alkynyl,cycloalkyl or cycloalkenyl; and

-   if R is

Y is CH, R^(c), R^(d), R^(e) and R^(fa) are all hydrogen, and L^(a) andL^(b) are both H or together form a pi-bond, then

if R^(a) is cyclohexyl, R^(b) is methyl, and R^(fb) is H then X is notphenyl, 4-biphenyl, 4-bromophenyl, 3-bromophenyl, 2-bromophenyl,4-tert-butylphenyl, 3-methoxyphenyl, 3,5-dimethoxyphenyl,3-(trifluoromethyl)phenyl, 4-(3,4-difluorophenyl)phenyl,4-(3-acetylphenyl)phenyl, 4-(4-methylthiophenyl)phenyl,4-(4-methoxyphenyl)phenyl, 4-(3-methoxyphenyl)phenyl,4-(2-methoxyphenyl)phenyl, 4-(3,5-dichlorophenyl)phenyl,4-(4-tert-butylphenyl)phenyl, 4-(3-ethoxyphenyl)phenyl,4-(3-chlorophenyl)phenyl, 4-(3-methylphenyl)phenyl,4-(4-methylphenyl)phenyl, 4-(2-methoxy-5-chlorophenyl)phenyl,4-(3-chloro-4-fluorophenyl)phenyl, 4-(4-trifluoromethoxyphenyl)phenyl,4-(3-trifluoromethoxyphenyl)phenyl, 4-(2,6-dimethoxyphenyl)phenyl,4-(4-dimethylaminophenyl)phenyl,

if R^(a) is cyclohexyl, R^(fb) is H and X is

then R^(b) is not methyl,

if R^(b) is methyl, R^(fb) is H and X is

then R^(a) is not cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl;

if R^(a) is cyclohexyl, R^(fb) is H and X is

then R^(b) is not methyl or tert-butyl;

if R^(a) is cyclohexyl, R^(b) is methyl, R^(fb) is H and X is

then R^(h) is not hydroxyl, (trimethylsilyl)ethoxymethyl-O, methoxy,O-benzyl, OCH₂CO₂Et, OC(O)CH₃, OC(O)Ph or OSO₂CH₃; and

-   if R^(a) is cyclohexyl, R^(b) is methyl, R^(fb) is H and X is

then R^(h) is not —CH═CHC(O)OMe, CH═CHC(O)OEt, CH═CHC(O)NMe₂,—CH═CHC(O)NH^(t)Bu, —CH═CHC(O)O^(t)Bu, CH═CHC(O)O^(i)Pr,—CH═CHC(O)OCH₂Ph, CH═CHC(O)OH, —CH═CHCH₂OMe, —CH═CHCH₂OEt or—CH═CHCH₂OPh.

In some embodiments, the compounds L_(c) and L_(d) are both H, and L^(a)and L^(b) together form a pi-bond.

Certain other disclosed compounds have the following general formula

With reference to the general formula, R¹ is selected from aryl,heteroaryl, heterocyclic, alkyl, alkenyl, cycloalkyl, cycloalkenyl orpolycyclic; R² is selected from alkyl, alkenyl, or cycloalkyl; Y isselected from N, N—O or C—R^(3d); R^(3a), R^(3b), R^(3c) and R^(3d) areeach independently selected from hydrogen, deuterium, halide, alkyl,alkenyl, alkoxy, alkylthio, amino, sulfonyl, aminosulfonyl,aminocarbonyl, acyl, hydroxyl or nitro; R^(4a) and R^(4b) are eachindependently selected from hydrogen, deuterium, halide or alkyl; L¹ andL² are independently selected from hydrogen, deuterium, alkyl,cycloalkyl, or together form a pi-bond; and R^(5a), R^(5b), R^(5c),R^(5d) and R^(5e) are each independently selected from hydrogen,deuterium, halide, alkyl, alkenyl, alkoxy, alkylthio, amino, sulfonyl,aminosulfonyl, aminocarbonyl, acyl, aryl, heteroaryl, cycloalkyl,heterocyclyl, hydroxyl or nitro; or any two adjacent groups selectedtogether form an aryl, heteroaryl, cycloalkyl or heterocyclic ring; andnone of R^(5a), R^(5b), R^(5c), R^(5d) or R^(5e) is —R^(x)-L^(x)-R^(x2),where R^(x) is selected from O, NR^(x3), sulfonyl or S; R^(x3) isselected from H, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, oraryl; L^(x) is selected from a bond, alkylene, alkenylene, alkynylene,cycloalkyl, cycloalkenyl, heterocyclic, aryl, heteroaryl orCR^(x4)R^(x5); R^(x4) and R^(x5) are each independently selected from H,D, halogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl,—C(O)OR^(x6), or —C(O)NR^(x6)R^(x7); R^(x6) and R^(x7) are eachindependently selected from H, alkyl, alkenyl, alkynyl, cycloalkyl orcycloalkenyl; R^(x2) is selected from —C(O)L^(x2)R^(x8) or a carboxylbioisostere; L^(x2) is a bond or NR^(x3); R^(x8) is H, alkyl, alkenyl,alkynyl, cycloalkyl, cycloalkenyl, —OR^(x9), N(R^(x9))₂, —C(O)R^(x9),—S(O)₂R^(x9), —C(O)OR^(x9), —S(O)₂N(R^(x9))₂ or —C(O)N(R^(x9))₂; andeach R^(x9) is independently selected from H, alkyl, alkenyl, alkynyl,cycloalkyl or cycloalkenyl. For certain embodiments where L¹ and L² areboth hydrogen or together form a pi-bond, Y is N or C-halogen; or R¹ ispolycyclic; or R⁴ is D, or R^(5a) is F, Cl or I; or R^(5d) and R^(5e)together form an aryl, heteroaryl, cycloalkyl or heterocyclic ring; orR^(5b) and R^(5c) together form an aryl, cycloalkyl, nitrogen-containingheterocyclic or nitrogen-containing heteroaryl ring; or any combinationthereof.

In some embodiments, Y is C—R^(3d), and R^(3d) or R^(5a) or both arehalogen, and in certain examples the halogen is fluorine. In otherembodiments, Y is N.

-   In certain embodiments, R¹ is polycyclic. Exemplary R¹ polycyclics    are selected from

or adamantyl. In other examples, the polycyclic is selected from[2.1.1], [2.2.1], [3.3.3], [4.3.1], [2.2.2], [4.2.2], [4.2.1], [4.3.2],[3.1.1], [3.2.1], [4.3.3], [3.3.2], [3.2.2], [3.3.1], [4.1.1], oradamantyl. In certain working embodiments, the polycyclic is

In some embodiments, R^(5c) is a nitrogen-containing heteroaryl ring,and the compound has a formula

where Z is selected from N, CH, or C-alkyl; R^(6a), R^(6c), R^(6d) andR^(6g) each is independently selected from H, D, halogen or alkyl; andR^(6h) is selected from H, D, alkyl, cycloalkyl, aryl or heteroaryl. Insome examples, Z is N, and/or R^(6a), R^(6c), R^(6d) and R^(6g) are allH. In particular embodiments, R^(6h) is methyl.

In other embodiments, R^(5c) comprises phenyl, leading to compoundshaving a formula

where R^(6a), R^(6b), R^(6c) and R^(6d) each is independently selectedfrom H, D, halogen or alkyl; G is a lone pair of electrons, or anoxygen; R^(6c) and R^(6f) each is independently selected from alkyl, Hor cycloalkyl; and

-   where R^(3d) or R^(5a) or both are halogen, or R⁴ is D, or R¹ is    polycyclic, or any combination thereof. In some examples, R^(6e) and    R^(6f) are both methyl.

In any of the above embodiments, R⁴ may be deuterium, and/or R² may bemethyl. In certain embodiments, R¹ is cyclohexyl.

In particular embodiments, the compound is selected from

Compositions comprising the disclosed compounds also are disclosed. Insome embodiments, the composition comprises a first disclosed compound,and an additional component, such as a pharmaceutically exceptableexcipient, an additional therapeutic compound, or a combination thereof.In certain examples, the additional therapeutic compound is a seconddisclosed compound. In some embodiments, the composition may include anenteric coating.

Also disclosed herein are embodiments of a method for treating orpreventing a metabolic disorder in a subject. Such methods can includeadministering to the subject a therapeutically effective amount of oneor more of the disclosed compounds, or one or more of the disclosedcompositions (such as 1, 2, 3, 4, or 5 of such compounds and/orcompositions). The compounds are substantially absorbed in thegastrointestinal tract, thereby activating FXR receptors in theintestines to treat or prevent a metabolic disorder in the subject. Themethod also may improve glucose and/or lipid homeostasis in the subject.In other embodiments, the method further includes administering to thesubject a statin, an insulin sensitizing drug, (such as sitagliptin,vildagliptin, saxagliptin, linagliptin, anaglptin, teneligliptin,alogliptin, gemiglptin, or dutoglpitin), meglitinide, sulfonylurea,peroxisome proliferator-activated receptor (alpha-glucosidase inhibitor,amylin agonist, dipeptidyl-peptidase 4 (DPP-4) inhibitor PPAR)-gammaagonist (e.g., a thiazolidinedione (TZD) [such as ioglitazone,rosiglitazone, rivoglitazone, or troglitazone], aleglitazar,farglitazar, muraglitazar, or tesaglitazar), a glucagon-like peptide(GLP) agonist, anti-inflammatory agent (e.g., oral corticosteroid),nicotinamide nibonucleoside and analogs thereof, or a combinationthereof.

In some examples, absorption of the compounds is substantially limitedto the intestines. In other examples, the compound substantiallyenhances FXR target gene expression in the intestines while notsubstantially enhancing FXR target gene expression in the liver orkidney.

In some embodiments, administering the compounds reduces or preventsdiet-induced weight gain and/or increases a metabolic rate in thesubject. Increasing the metabolic rate may include enhancing oxidativephosphorylation in the subject.

In some embodiments, administering the compounds results in nosubstantial change in food intake and/or fat consumption in the subject,and/or no substantial change in appetite in the subject. Administeringthe compounds can protect against diet-induced weight gain, reduceinflammation, enhance thermogenesis, enhance insulin sensitivity in theliver, reduce hepatic steatosis, promote browning of white adiposetissue (WAT), promote activation of brown adipose tissue (BAT), decreaseblood glucose, increase weight loss, or any combination thereof. Inparticular embodiments, administering the compounds enhances insulinsensitivity in the liver and promotes BAT activation.

Exemplary metabolic disorders include but are not limited to: obesity(such as a BMI of greater than 25, at least 30, at least 35, or at least40, such as 25 to 30, 35 to 40, or over 40), diabetes, insulinresistance, dyslipidemia (such as an elevated serum lipids and/ortriglycerides, such as a serum LDL of at least 100 mg/dL, such as atleast 130 mg/dL, at least 160 mg/dL or at least 200 mg/dL, such as 100to 129 mg/dL, 130 to 159 mg/dL, 160 to 199 mg/dL or greater than 200mg/dL, and/or such as a serum triglyceride of at least of at least 151mg/dL, such as at least 200 mg/dL, or at least 500 mg/dL, such as 151 to199 mg/dL, 200 to 499 mg/dL or greater than 499 mg/dL) or anycombination thereof. In particular examples, the metabolic disorder isnon-insulin dependent diabetes mellitus.

Embodiments of a method for treating or preventing inflammation in anintestinal region of a subject are also disclosed. Administering to asubject a therapeutically effective amount of one or more of thedisclosed compounds, or one or more of the disclosed compositions, suchas 1, 2, 3, 4, or 5 of such compounds and/or compositions, activates FXRreceptors in the intestines, thereby treating or substantiallypreventing inflammation in the intestinal region of the subject. In someembodiments, the method further includes administering a therapeuticallyeffective amount of an antibiotic (such as metronidazole, vancomycin,and/or fidaxomicin) to the subject, such as to treat or substantiallyprevent inflammation associated with pseudomembranous colitis in thesubject. In other embodiments, the method comprises administering to thesubject a therapeutically effective amount of an oral corticosteroidand/or other anti-inflammatory or immunomodulatory therapy incombination with the compound, and/or in combination with an antibiotic.

Intestinal inflammation may be associated with a clinical conditionselected from necrotizing enterocolitis, gastritis, ulcerative colitis,Crohn's disease, inflammatory bowel disease, irritable bowel syndrome,gastroenteritis, radiation induced enteritis, pseudomembranous colitis,chemotherapy induced enteritis, gastro-esophageal reflux disease (GERD),peptic ulcer, non-ulcer dyspepsia (NUD), celiac disease, intestinalceliac disease, post-surgical inflammation, gastric carcinogenesis,infectious colitis, or any combination thereof. In certain examples, theone or more FXR target genes comprises IBABP, OSTα, Per1, FGF15, FGF19,or combinations thereof.

Embodiments of a method for treating or preventing cholestatic disordersin subject (such as an adult or pediatric subject) are also disclosed.Administering to a subject a therapeutically effective amount of one ormore of the disclosed compounds, or one or more of the disclosedcompositions, such as 1, 2, 3, 4, or 5 of such compounds, can be used totreat or prevent a cholestatic disorder in subject. Cholestasis is acondition where bile cannot flow (or flow is significantly reduced) fromthe liver to the duodenum, for example due to a mechanical blockage(e.g., gallstone, malignancy, or congenital defect), or as a result of adefect in bile formation (e.g., due to a genetic defect, side effect ofmedication). Examples of such disorders include, but are not limited to,primary biliary cirrhosis (PBC), primary sclerosing cholangitis (PSC),overlap syndrome (PBC plus autoimmune hepatitis), cholestasis resultingfrom a drug (e.g., one or more of androgen, birth control pills, goldsalts, nitrofurantoin, anabolic steroids, chlorpromazine,prochlorperazine, sulindac, cimetidine, estrogen, statins, andantibiotics such as TMP/SMX, flucoxacillin and erythromycin),drug-induced cholestatic hepatitis, total parenteral nutrition(TPN)-induced cholestasis, ICU/sepsis-related cholestasis, obstetriccholestasis, graft vs. host disease, prolonged cholestasis due tohepatitis A, B or C infection, cholestasis due to cystic fibrosis,alcoholic hepatitis, progressive familial intrahepatic cholestasis(PFIC) syndromes, Alagille syndrome, biliary atresia, or any combinationthereof. In some embodiments, the method further includes administeringa therapeutically effective amount of another therapeutic agent (such asursodeoxycholic acid, phenobarbital, methotrexate, fat-soluble vitamins,or combinations thereof) to the subject, such as to treat orsubstantially prevent one or more cholestatic disorders in the subject.

Embodiments of a method for treating or preventing intestinalpermeability conditions in subject are also disclosed. Administering toa subject a therapeutically effective amount of one or more of thedisclosed compounds, or one or more of the disclosed compositions, suchas 1, 2, 3, 4, or 5 of such compounds, can be used to treat or preventan intestinal permeability condition in subject. Intestinal permeabilityis a condition where the gut wall exhibits excessive permeability (whichsome in the field call leaky gut syndrome). Examples of such disordersinclude, but are not limited to, Crohn's disease, ulcerative colitis,infectious colitis, celiac disease, type 1 diabetes, inflammatory boweldisease, irritable bowel syndrome, or any combination thereof. In someembodiments, the method further includes administering a therapeuticallyeffective amount of another therapeutic agent (such as glutamine,prebiotics, probiotics, Escherichia coli Nissle 1917, or combinationsthereof) to the subject, such as to treat or substantially prevent oneor more intestinal permeability disorders in the subject.

Embodiments of a method for treating or preventing disorder that causesor results from an altered intestinal microbiome in subject are alsodisclosed. Administering to a subject a therapeutically effective amountof one or more of the disclosed compounds, or one or more of thedisclosed compositions, such as 1, 2, 3, 4, or 5 of such compounds, canbe used to treat or prevent a disorder resulting altered intestinalmicrobiome in subject. An altered intestinal microbiome is a conditionwhere the abundance and/or types of bacteria (such as Bacteriodes. E.coli, Lactobacillus, and Bifidobacteria species) and other microbes(such as yeast) in the intestine are abnormal. Examples of disordersthat can have an altered gut microbiome include, but are not limited to,celiac disease, the intestinal permeability conditions described herein,the intestinal inflammation disorders described herein, alcoholichepatitis, necrotizing enterocolitis, Crohn's disease, ulcerativecolitis, intestinal lesions (such as those in a cystic fibrosispatient), cirrhosis, or any combination thereof. In some embodiments,the method further includes administering a therapeutically effectiveamount of another therapeutic agent (such as a fecal microbiotatransplant, immunosuppressant, antibiotic, mesalamine, steroid, altereddiet, or combinations thereof) to the subject, such as to treat orsubstantially prevent one or more disorders resulting from or thatcauses an altered intestinal microbiome in the subject.

Embodiments of a method for treating an inborn error of metabolism insubject are also disclosed. Administering to a subject a therapeuticallyeffective amount of one or more of the disclosed compounds, or one ormore of the disclosed compositions, such as 1, 2, 3, 4, or 5 of suchcompounds, can be used to treat or prevent an inborn error of metabolismin subject. An inborn error of metabolism is a genetic conditionresulting in accumulation of substance which interfere with normalfunction or the reduced ability to synthesize essential compounds, (suchas a reduction in bile acid production, lipid production, or lipidstorage). One example of an inborn error of metabolism iscerebrotendinous xanthomatosis (CTX). In some embodiments, the methodfurther includes administering a therapeutically effective amount ofanother therapeutic agent (such as chenodeoxycholic acid (CDCA), anHMG-CoA reductase inhibitor (“statins” such as simvastatin) orcombinations thereof) to the subject, such as to treat an inborn errorof metabolism in the subject.

Embodiments of a method for treating or preventing a bile disorder insubject are also disclosed. Administering to a subject a therapeuticallyeffective amount of one or more of the disclosed compounds, or one ormore of the disclosed compositions, such as 1, 2, 3, 4, or 5 of suchcompounds, can be used to treat or prevent a bile disorder in subject.Bile disorders include mechanical biliary obstructions, disorders thatresult from bile acid malabsorption, and bile acid synthesis disorders.Examples of bile disorders that can be treated with the disclosedcompounds include, but are not limited to, benign biliary stricture,malignant biliary obstruction, bile acid diarrhea, or any combinationthereof. In some embodiments, the method further includes administeringa therapeutically effective amount of another therapeutic agent (such asbile acid sequestrant, cholestyramine, colestipol, farnesoid X receptoragonist (such as obeticholic acid), or combinations thereof) to thesubject, such as to treat or prevent a bile disorder in the subject.

Embodiments of a method for treating or preventing a malabsorptiondisorder (e.g., intestinal malabsorption), such as short bowel syndrome(or symptoms arising from such, such as diarrhea, steatorhea,malnutrition, fatigue, vitamin deficiency), environmental enteropathy,or tropical sprue, in subject are also disclosed. Administering to asubject a therapeutically effective amount of one or more of thedisclosed compounds, or one or more of the disclosed compositions, suchas 1, 2, 3, 4, or 5 of such compounds, can be used to treat amalabsorption disorder in subject. Short bowel syndrome is amalabsorption disorder causes by surgical removal of the small intestineor dysfunction of a large segment of bowel. Short bowel syndrome can becaused by a birth defect, Crohn's disease, volvulus, tumors, injury,necrotizing enterocolitis, or surgery. Environmental enteropathy is amalabsorption disease believed to be due to frequent intestinalinfections, which can result in chronic malnutrition and growthstunting. Tropical sprue is a malabsorption disease found in tropicalregions, with abnormal flattening of the villi and inflammation of thesmall intestine. In some embodiments, the method further includesadministering a therapeutically effective amount of another therapeuticagent (such as an anti-diarrheal medicine such as loperamide or codeine,vitamin supplement (such as B₁₂ and folic acid), mineral supplement, Lglutamine, proton pump inhibitors, lactase, tedulutide (a glucagon-likepeptide-2 analog), total parenteral nutrition, antibiotic (e.g.,tetracycline or sulfamethoxazole/trimethoprim) or combinations thereof)to the subject, such as to treat or prevent a malabsorption disorder inthe subject.

Embodiments of a method for treating or preventing a cell proliferationdisease (e.g., cancer, such as adenocarcinoma, such as cancer of thecolon, jejunum, and/or ileum), for example in an intestinal region of asubject, are also disclosed. Administering to a subject atherapeutically effective amount of one or more of the disclosedcompounds, or one or more of the disclosed compositions, such as 1, 2,3, 4, or 5 of such compounds and/or compositions, activates FXRreceptors in the intestines, thereby treating or substantiallypreventing a cell proliferation disease, for example in the intestinalregion of the subject. In some embodiments, the method further includesadministering a therapeutically effective amount of another therapeuticagent, (such as a chemotherapeutic, a biologic, a radiotherapeutic, orcombinations thereof) to the subject, such as to treat or substantiallyprevent a cell proliferation disease in the subject.

Embodiments of a method for treating or preventing alcoholic liverdisease (e.g., fatty liver (steatosis), cirrhosis, alcoholic hepatitis),nonalcoholic steatohepatitis (NASH), or nonalcoholic fatty liver disease(NAFLD), in a subject, are also disclosed. Administering to a subject atherapeutically effective amount of one or more of the disclosedcompounds, or one or more of the disclosed compositions, such as 1, 2,3, 4, or 5 of such compounds and/or compositions, can treat orsubstantially preventing alcoholic liver disease, NASH, or NAFLD. Insome embodiments, the method further includes administering atherapeutically effective amount of another therapeutic agent, (such asa corticosteroid, anti-tumor necrosis factor (TNF) or combinationsthereof) to the subject, such as to treat or substantially preventalcoholic liver disease, NASH, or NAFLD in the subject.

In any of the above embodiments, the method may increase HSLphosphorylation and β3-adrenergic receptor expression (such as anincrease of at least 20%, at least 25%, at least 30%, at least 40%, atleast 50%, at least 75%, or at least 100%). Additionally, the serumconcentration of the compound in the subject may remain below its EC₅₀following administration of the compound.

Also disclosed herein are embodiments of a method for making thedisclosed compounds. In some embodiments, the method comprises reactingan aldehyde with a first amine to form an imine, reacting the imine witha reducing agent to form a second amine, and reacting the second aminewith an activated carboxylic acid derivative or a carboxylic acid toform an amide. In certain embodiments, the method further comprisingreacting the aldehyde with a boronic acid, and/or reacting the amidewith a vinyl ester. In other embodiments, the method further comprisesreacting the first amine with a vinyl ester, and/or reacting the amidewith a boronic acid. The reducing agent may be a deuterated reducingagent to produce compounds comprising deuterium.

The foregoing and other objects and features of the disclosure willbecome more apparent from the following detailed description, whichproceeds with reference to the accompanying figures.

BRIEF DESCRIPTION OF THE DRAWINGS

This application contains at least one drawing executed in color. Copiesof any patent(s) issuing from this application, or patent applicationpublication(s), with color drawing(s) will be provided by the Officeupon request and payment of the necessary fee.

FIGS. 1A-1C are a comparative expression chart and two bar charts,respectively, illustrating increased levels of FXR target geneexpression in the intestine relative to expression in the liver andkidney. 8 week-old C57BL/6J mice were treated with vehicle or fexaramine(100 mg/kg) via oral (PO) or intraperitoneal (IP) injection for threedays (FIGS. 1A-1B) or five days (FIG. 1C).

FIG. 1A shows FXR target SHP gene expression in FXR abundant tissuesincluding liver, kidney and intestine from 8 week-old mice that weretreated with vehicle or fexaramine (100 mg/kg) via oral (PO) orintraperitoneal (IP) injection for three days. FXR target geneexpression was analyzed by qPCR. Gene expression was normalized againsta vehicle-treated group.

FIG. 1B shows that PO administration of fexaramine (solid bars), but notvehicle (open bars), substantially enhances FXR target gene expressionin the intestine, and not in the liver or kidney.

FIG. 1C shows that IP injection of fexaramine increases FXR target geneexpression in the liver and kidney, in addition to the intestines. Datarepresent the mean ±SD. Statistical analysis was performed with theStudent's t test. *p<0.05, **p<0.01

FIG. 1D is a schematic diagram illustrating an experimental procedureused to evaluate fexaramine, where mice were treated with vehicle orfexaramine (100 mg/kg) via PO or IP injection, and LC/MS quantificationof serum fexaramine was conducted five days later.

FIG. 1E is a bar chart illustrating serum fexaramine concentrationsafter administration as described in FIG. 1D. Data represent mean values±STD. Statistical analysis was performed with the Student's t test(*p<0.05, **p<0.01).

FIG. 1F is a bar chart illustrating that orally delivered fexaramine isintestinally-restricted. Mice received vehicle or Fexaramine (100 mg/kg)via per os (PO) or intraperitoneal (IP) injection for 5 days. Expressionof the FXR target gene SHP after PO or IP injection in selected tissuesis shown.

FIGS. 2A-2G are graphs illustrating the reduction of diet-inducedobesity and improvement in metabolic homeostasis with fexaramine Micewere fed a high fat diet (HFD) for 14 weeks and then administered dailyoral injections of vehicle (open boxes) or fexaramine (100 mg/kg) (solidboxes) for 5 weeks with HFD. Data represent the mean ±STD. Statisticalanalysis was performed with the Student's t test (*p<0.05, **p<0.01).

FIG. 2A is a line chart illustrating changes in body weight of mice feda high fat diet (HFD) for 14 weeks and then administered daily oralinjections of vehicle (open boxes) or fexaramine (100 mg/kg) (solidboxes) for 5 weeks with HFD. n=8 per group.

FIG. 2B shows mice body weight composition by MRI at the completion ofthe study.

FIG. 2C shows the wet weight of inguinal fat (iWAT), gonadal fat (gWAT),mesenteric fat (mWAT), liver, kidney, heart and spleen at the completionof the study.

FIG. 2D shows the serum levels (samples were collected after 8hours-fasting for parameter analysis) of insulin, cholesterol, leptin,resistin and triglycerides.

FIG. 2E shows the serum levels of cytokines at the completion of thestudy.

FIG. 2F is a line graph representing glucose tolerance testing (GTT),which revealed that fexaramine treatment improved glucose clearance.

FIG. 2G is a line graph representing insulin tolerance testing (ITT),which showed that fexaramine treatment improved insulin sensitivity.

FIGS. 3A-3D are line graphs and a bar graph showing the effects offexaramine administration in normal chow-fed mice. The mice were treatedwith vehicle or fexaramine (100 mg/kg) via PO for 5 weeks. Datarepresent the mean ±STD. Statistical analysis as performed with theStudent's t test (*p<0.05, **p<0.01).

FIG. 3A is a line graph showing hourly composite carbon dioxideproduction.

FIG. 3B is a line graph showing hourly composite oxygen consumption.

FIG. 3C is a glucose tolerance test.

FIG. 3D is a bar graph showing core body temperature.

FIG. 4A is a line graph showing the effects of fexaramine at variousdosage levels on the body weight of mice fed a HFD for 14 weeks and thenadministered daily oral injections of vehicle or fexaramine (10, 50 or100 mg/kg) for 5 weeks with HFD. Data represent the mean ±STD.Statistical analysis was performed with the Student's t test (*p<0.05,**p<0.01).

FIG. 4B is a set of digital images showing histological analysis of theileum and colon following treatment with fexaramine or vehicle. Micewere fed on HFD for 14 weeks, and then administered daily oralinjections of vehicle or fexaramine (100 mg/kg) for 5 weeks with HFD.

FIG. 4C is a line graph showingn glucose tolerance tests in mice fed aHFD for 14 weeks and then administered daily oral injections of vehicleor fexaramine (10, 50 or 100 mg/kg) for 5 weeks with HFD. Data representthe mean ±STD. Statistical analysis was performed with the Student's ttest (*p<0.05, **p<0.01).

FIG. 4D is a line graph showintfasting glucose levels in 14 week HFD-fedmice treated with vehicle or fexaramine (100 mg/kg/day os for 5 week).Data represent the mean ±STD. Statistical analysis was performed withthe Student's t test (*p<0.05, **p<0.01).

FIGS. 5A-5I show that FXR is required for fexaramine's effects (A) Bodyweights, (B) glucose tolerance test, (C) insulin tolerance test, (D)oxygen consumption, (E) carbon dioxide production, (F) core bodytemperature, (G) brown adipose tissue gene expression, (H) liver geneexpression, and (I) FXR target gene expressions in ileum of 14 week HFDfed FXR-null mice treated with vehicle or fexaramine (100 mg/kg) for 5week with HFD. Data represent the mean ±SD. Statistical analysis wasperformed with the Student's t test. *p<0.05, **p<0.01.

FIGS. 6A-6J demonstrate that fexaramine increases OXPHOS to enhancemetabolic rate in brown adipose tissue. Mice were fed HFD for 14 weeksand then administered vehicle or fexaramine (100 mg/kg) daily by oraladministration for 5 weeks with HFD. Data represent the mean ±STD.Statistical analysis was performed with the Student's t test (*p<0.05,**p<0.01).

FIG. 6A is a bar chart showing daily food intake during the first weektreatment.

FIG. 6B is a line chart showing carbon dioxide production.

FIG. 6C is a line chart showing oxygen consumption.

FIG. 6D is a bar chart showing daytime and nighttime cumulativeambulatory counts.

FIG. 6E is a bar chart showing core body temperature.

FIG. 6F shows hematoxyin and eosin staining of brown adipose tissue(BAT) for histological analysis.

FIG. 6G is a bar chart showing relative gene expression of nuclearreceptors and other genes encoding proteins involved in mitochondrialbiogenesis, glucose transport and FA oxidation in BAT.

FIG. 6H is a set of digital images of gel electrophoreses showingprotein expression levels of total and phosphorylated p38 in BAT. RalAlevels are shown as a loading control.

FIG. 6I is a bar chart showing the relative levels of phosphorylated p38in BAT after vehicle (open bar) or Fexaramine administration (solidbar).

FIG. 6J is a chart showing changes in relative expression of OXPHOSgenes based on RNA-sequencing transcriptomic analysis in inguinal fat(iWAT), gonadal fat (gWAT) and brown fat (BAT) after vehicle orfexaramine treatment.

FIG. 6K is a heatmap depiction of changes in genes involved in chemokineand cytokine signaling in BAT after vehicle or fexaramine treatment.

FIG. 6L is a bar graph showing PKA activity in BAT. Data represent themean ±SD. Statistical analysis was performed with the Student's t test.*p<0.05, **p<0.01.

FIG. 6M is a bar chart showing the effect of fexaramine on respiratoryexchange ratio (RER). Mice were fed on HFD for 14 weeks, and thenadministered daily oral injections of vehicle or fexaramine (100 mg/kg)for 5 weeks with HFD. No changes were observed in respiratory exchangeratio by fexaramine treatment.

FIG. 6N is a bar graph showing the effect of fexaramine administrationon serum lactate concentrations. Mice were fed on HFD for 14 weeks, andthen administered daily oral injections of vehicle or fexaramine (100mg/kg) for 5 weeks with HFD. Serum lactate levels were found to besignificantly decreased with fexaramine treatment. Data represent themean ±STD. Statistical analysis was performed with the Student's t test(*p<0.05, **p<0.01).

FIGS. 7A-7H show a comparative expression chart and bar chartsillustrating that fexaramine increased endogenous FGF15 signaling andchanges in BA composition. Mice were fed HFD for 14 weeks and thenadministered daily oral injections of vehicle or fexaramine (100 mg/kg)for 5 weeks with HFD. In the bar graphs, open bars represent vehicletreatment and solid bars represent fexaramine treatment, and datarepresent the mean ±STD. Statistical analysis was performed with theStudent's t test (*p<0.05, **p<0.01).

FIG. 7A is a heatmap depicting changes in expression of ileal FXR targetgenes following PO fexaramine administration.

FIG. 7B is a bar chart showing FGF15 protein levels from ileal extract.

FIG. 7C is a bar chart showing FGF15 protein levels in the serum.

FIG. 7D is a bar chart showing changes in the expression of hepaticgenes involved in bile acid metabolism.

FIG. 7E is a bar chart showing total serum bile acid (BA) levels.

FIG. 7F is a bar chart showing composition ratios of bile acids. Theratio of unconjugated to conjugated cholic acid was remarkably increasedby fexaramine

FIG. 7G is a bar chart showing changes in intestinal permeability.

FIG. 7H is a bar chart showing changes in expression of intestinal genesinvolved in mucosal defense.

FIG. 8 is a bar graph showing hepatic Cyp7a1 levels determined by ELISA.Data represent the mean ±SD. Statistical analysis was performed with theStudent's t test. *p<0.05, **p<0.01.

FIG. 9 is a bar graph showing that fexaramine fails to activate TGR5.HEK293 cells were transfected with expression vectors for cAMP-responseelement luciferase, β-galactosidase and human TGR5. 24 hours aftertransfection, cells were treated with fexaramine or INT-777 (a TGR5agonist).

FIGS. 10A-10F show that systemic TGR5 activation is required to affectglucose homeostasis. HFD-fed mice were treated with vehicle, theintestinally-restricted TGR5 ligand L755-0379 (A, L755, 100 mg/kg, EC50300 nM) or the systemic ligand RO5527239 (B, RO, 100 mg/kg. EC50 70 nM)via per os for 14 days. C, Plasma L755 concentrations in portal and tailveins after PO administration. D, Body weight curve. E, Glucosetolerance test. F, Serum insulin levels after a glucose challenge. Datarepresent the mean ±SD. Statistical analysis was performed with theStudent's t test. *p<0.05, **p<0.01.

FIGS. 11A-11N show that TGR5 is required for a subset of fexaramine'seffects. HFD-fed TGR5-null mice were treated with vehicle or fexaramine(100 mg/kg os daily for 5 weeks with HFD, n=10). (A) Heal FXR targetgene expressions (B) Serum BA levels (C) Fasting glucose levels (D)Glucose tolerance test (E) Core body temperature (F) Oxygen consumptionrate (G) Carbon dioxide production (H) Gene expression in BAT (I) Bodyweight curve (J) Body composition by MRI (K) Insulin Tolerance Test (L)Hepatic gene expression (M) Hepatic TG levels (N) and Gene expression insoleus of TGR5 knockout mice with and without fexaramine treatment. Datarepresent the mean ±SD. Statistical analysis was performed with theStudent's t test. *p<0.05, **p<0.01.

FIGS. 12A-12H demonstrate that fexaramine reduces inflammation andincreases lipolysis in adipose tissues. Mice were fed on HFD for 14weeks and subsequently subjected to daily PO injection of vehicle orfexaramine (100 mg/kg) for 5 weeks with HFD. In the bar graphs, openbars are vehicle, solid bars of fexaramine, and data represent the mean±STD. Statistical analysis was performed with the Student's t test(*p<0.05, **p<0.01).

FIG. 12A shows histological sections of mesenteric white adipose tissuesfrom vehicle and fexaramine-treated mice.

FIG. 12B is a set of photographs of gel electrophoreses showing proteinexpression levels of TBK1, and total and phosphorylated IKKε and S6K, ingonadal adipose tissues (gWAT) from vehicle or fexaramine-treated mice.

FIG. 12C is a bar chart showing relative gene expression levels ofβ-3-adrenergic receptor and various cytokines in gonadal adipose tissue.

FIG. 12D is a set of photographs of gel electrophoreses showing proteinexpression levels of total and phosphorylated HSL (p-HSL) and p65 ingonadal and inguinal adipose tissues.

FIG. 12E is a bar chart showing serum levels of catecholamines, invehicle or fexaramine-treated mice.

FIG. 12F is a bar chart showing serum glycerol levels, in vehicle orfexaramine-treated mice. Isoproterenol (1 μg/kg) was injected at 0minutes and free glycerol levels were measured at the indicated timepoints.

FIG. 12G is a bar chart showing serum levels of free fatty acids invehicle or fexaramine-treated mice. Data represent the mean ±STD.Statistical analysis was performed with the Student's t test (*p<0.05,**p<0.01).

FIG. 12H shows UCP1 staining of brown fat-like cells in inguinal adiposetissues (iWAT) from vehicle or fexaramine-treated mice(Magnification:100×).

FIGS. 12I and 12J show that fexaramine enhances OXPHOS in iWAT. Mice feda HFD for 14 weeks were maintained on a HFD and treated with vehicle orfexaramine (100 mg/kg/day os for 5 week). (I) Changes in genesassociated with the browning of adipose tissue and (J) oxygenconsumption rate of the stromal vascular fraction (SVF) from inguinalfat (iWAT). Data represent the mean ±SD. Statistical analysis wasperformed with the Student's t test. *p<0.05, **p<0.01.

FIG. 13 is a set of digital images of gel electrophoreses (Westernblots) showing the level of expression of various proteins in gonadalwhite adipose tissue (gWAT). Mice fed a HFD for 14 weeks were maintainedon a HFD and treated with vehicle or fexaramine (50 mg or 100 mg/kg/dayos for 5 week).

FIG. 14 is a bar chart showing that fexaramine reduces brown adiposetissue (BAT) inflammation. Mice fed a HFD for 14 weeks were maintainedon a HFD and treated with vehicle or fexaramine (100 mg/kg/day os for 5week). Expression of inflammatory cytokines in BAT. Data represent themean ±SD. Statistical analysis was performed with the Student's t test.*p<0.05, **p<0.01.

FIGS. 15A-15H are a set of histology stains and bar charts demonstratingthat fexaramine induced less weight gain and improved glucosehomeostasis relative to mice that did not receive fexaramine. Mice werefed HFD for 14 weeks and then subjected to daily PO injection of vehicleor fexaramine (100 mg/kg) for 5 weeks with HFD.

FIG. 15A is a bar chart showing basal hepatic glucose production (HGP).

FIG. 15B is a bar chart showing glucose disposal rate (GDR).

FIG. 15C is a bar chart showing percentage free fatty acid (FFA)suppression by insulin.

FIG. 15D is a bar chart showing HGP suppression by insulin, as measuredby hyperinsulinemic-euglycemic clamps.

FIG. 15E shows hematoxylin and eosin staining for liver histology.

FIG. 15F is a bar chart showing triglyceride levels in the liver.

FIG. 15G is a bar chart showing hepatic gene expression levels for genesinvolved in gluconeogenesis and lipogenesis.

FIG. 15H is a bar chart showing serum levels of alanine aminotransferase(ALT).

FIGS. 15I-15K are a line graph and two bar graphs showing the effect offexaramine treatment on body weight, insulin-stimulated GDR, and fastinginsulin levels. Mice were fed HFD for 14 weeks, and then administereddaily oral injections of vehicle or fexaramine (100 mg/kg) for 3 weekswith HFD. The mice treated with fexaramine were initially heavier (by2-3 grams). Three weeks after treatment, a clamp study was performed onthe mice. Data represent the mean ±STD. Statistical analysis wasperformed with the Student's t test (*p<0.05, **p<0.01).

FIG. 15I is a line graph showing the changes in body weight for the twogroups of mice.

FIG. 15J is a bar chart showing the insulin-stimulated GDR (IS-GDR).

FIG. 15K is a bar chart showing the fasting insulin levels.

FIGS. 16A, 16B and 16C are graphs of percentage activation of FXR versusthe log value of concentration for duplicate runs of NSSK00024,fexaramine and DMSO.

FIGS. 17A, 17B and 17C are graphs of percentage activation of FXR versusthe log value of concentration for duplicate runs of NSSK00027,fexaramine and DMSO.

FIGS. 18A, 18B and 18C are graphs of percentage activation of FXR versusthe log value of concentration for duplicate runs of NSSK00089,fexaramine and DMSO.

FIGS. 19A, 19B and 19C are graphs of percentage activation of FXR versusthe log value of concentration for duplicate runs of NSSK00096,fexaramine and DMSO.

FIGS. 20A, 20B and 20C are graphs of percentage activation of FXR versusthe log value of concentration for duplicate runs of NSSK00110,fexaramine and DMSO.

FIG. 21 is a graph of percentage activation of FXR versus the log valueof concentration for NSSK00001, fexaramine and DMSO.

FIG. 22 is a graph of percentage activation of FXR versus the log valueof concentration for NSSK00006, fexaramine and DMSO.

FIG. 23 is a graph of percentage activation of FXR versus the log valueof concentration for NSSK00026, fexaramine and DMSO.

FIG. 24 is a graph of percentage activation of FXR versus the log valueof concentration for NSSK00039, fexaramine and DMSO.

FIG. 25 is a graph of percentage activation of FXR versus the log valueof concentration for NSSK00048, fexaramine and DMSO.

FIG. 26 is a graph of percentage activation of FXR versus the log valueof concentration for NSSK00057, fexaramine and DMSO.

FIG. 27 is a graph of percentage activation of FXR versus the log valueof concentration for NSSK00086, fexaramine and DMSO.

FIGS. 28A and 28B are graphs of percentage activity versus log value ofthe molar concentration for duplicate runs of deuterated fexaramine,fexaramine and DMSO.

FIG. 29A-H show the effect of fexaramine and selectively-deuteratedfexaramine analogs in vivo. (A) Structures of Fexaramine and analogsSALK24 (NSSK00024) and SALK110 (NSSK00110) indicating positions ofselective deuteration. (B) Body weights of mice during course of drugtreatment. (C) Core body temperature of mice before (Day 0) and after(day 14) treatment with the indicated FXR analogs. (D) Changes in thefasting glucose levels of ob/ob mice after treatment with the indicatedFex analogs for 1 and 2 weeks. (E) Glucose tolerance test (GTT)performed on ob/ob mice after 2 weeks treatment with the indicatedanalogs. (F) Fasting insulin levels in ob/ob mice after 2 weekstreatment with the indicated analogs. (G) Insulin secretion, measuredduring a GTT assay, in ob/ob mice after 2 weeks treatment with theindicated analogs. (H) Glucagon-like peptide-1 (GLP1) secretion,measured during a GTT assay, in ob/ob mice after 2 weeks treatment withthe indicated analogs.

FIG. 30 is a bar graph showing expression of several genes in the liverafter treatment with Fex-D or Salk110, as measured by QPCR. Datarepresent the mean ±STD. Statistical analysis was performed with theStudent's t test (*p<0.05, **p<0.01).

FIG. 31 is a heatmap comparing expression changes in selected genesinduced by fexaramine analogs.

FIG. 32 is a table showing the relative transport rates of fexaramineand fexaramine analogs provided herein (NSSK00024, NSSK00027, NSSK00089,NSSK00096, and NSSK00110), as well as controls, in Caco2 cells.

FIGS. 33A-33C show fexaramine-treated APC^(min) mice are resistant tocachexia. (A) Body weight measurements of vehicle or Fex-treatedAPC^(min) mice. (B) Normalized body weight changes of vehicle or Fextreated APC^(min) mice. (C) Survival curves of vehicle and Fex-treatedAPC^(min) mice.

FIGS. 34A-34D show fexaramine-treated APC^(min) mice have reduced tumorburden. (A) Total tumor burden of vehicle or Fex treated APC^(min) miceafter 23 weeks. (B) Duodenal tumor burden of vehicle or Fex treatedAPC^(min) mice after 23 weeks. (C) Jejunal tumor burden of vehicle orFex treated APC^(min) mice after 23 weeks. (D) Ileal tumor burden ofvehicle or Fex treated APC^(min) mice after 23 weeks.

FIGS. 35A-35D show fexaramine treatment of APC^(min) mice reduces tumorsize and distribution. (A) Tumor size distribution in the duodenum invehicle and Fex treated APC^(min) mice after 23 weeks. (B) Tumor sizedistribution in the jejunum in vehicle and Fex treated APC^(min) miceafter 23 weeks. (C) Tumor size distribution in the lieum in vehicle andFex treated APC^(min) mice after 23 weeks. (D) Tumor size distributionthroughout the intestine in vehicle and Fex treated APC^(min) mice after23 weeks.

FIG. 36 is a digital image of serum from APC^(min) mice, showing thatfexaramine-treatment reduces circulating triglycerides.

FIG. 37 is a digital image showing duodenum paraformaldehyde fixedintestinal sections of APC^(min) mice with reduced tumor sizeRepresentative images of fixed duodenum tissue of vehicle or Fex treatedAPC^(min) mice after 23 weeks treatment. White triangles point toidentified tumors and showed a reduction in tumor size and number in Fexcompared to Vehicle treated mice.

FIG. 38 is a digital image showing jejunum paraformaldehyde fixedintestinal sections of APC^(min) mice showed reduced tumor size.Representative images of fixed jejunum tissue of vehicle or Fex treatedAPC^(min) mice after 23 weeks treatment. White triangles point toidentified tumors and showed reduction in tumor size and number in Fexcompared to Vehicle treated mice.

FIG. 39 is a digital image showing ileum paraformaldehyde fixedintestinal sections of APC^(min) mice showed reduced tumor size.Representative images of fixed ileum tissue from vehicle or Fex treatedAPC^(min) mice after 23 weeks treatment. White triangles point toidentified tumors and showed reduction in tumor size and number in Fexcompared to Vehicle treated mice.

FIG. 40 is a digital image showing colon paraformaldehyde fixedintestinal sections of APC^(min) mice showed reduced tumor size.Representative image of fixed colon tissue of vehicle or Fex treatedAPC^(min) mice after 23 weeks treatment. White triangles point toidentified tumors. No tumors were observed in Fex treated mice comparedwith vehicle treated which have tumors.

FIG. 41 is a graph showing the effects of modulating FXR activity on theoxidative metabolism of intestinal L cells. L cells were treated withthe FXR agonist fexaramine, the FXR antagonist Guggulsterone, or acombination of both, prior to measurement of their oxygen consumptionrate (OCR) in a Seahorse analyzer.

FIGS. 42A-42C demonstrate the metagenomics and metabolomics in alcoholicliver disease. (A) schematic drawing showing that the activity of theenzyme choloylglycine hydrolase, responsible for the deconjugation ofbile acids, is increased in alcohol liver disease (ALD). (B) a bar graphshowing the levels of conjugated and unconjugated bile acids in theplasma of C57BL/6J mice after intragastric feeding of an isocaloric dietor ethanol for 3 weeks and (C) a bar graph showing the levels ofconjugated and unconjugated bile acids in the liver of mice afterintragastric feeding of an isocaloric diet or ethanol for 3 weeks.

FIGS. 43A and 43B are a (A) digital image of a western blot of Cyp7a1protein levels in the livers of mice following 3 weeks intragastricfeeding of an isocaloric diet (control) or ethanol (tubulin is providedas a protein loading control) and (B) a bar graph quantifying Cyp7a1protein levels, as measured by Western blot.

FIGS. 44A-44C are a (A) digital image of histology images of the liverand (B) and (C) bar graphs showing that administration of fexaramine canprotect the liver from alcoholic liver disease, for example bydecreasing fat in the liver, liver enzyme ALT and triglycerides (TG).C57BL/6J mice, fed an isocaloric diet or ethanol through continuousintragastric feeding for 3 weeks, were co-administered Fexaramine (100mg/kg/day oral gavage) or vehicle. (A) Histological liver sections fromethanol fed mice after vehicle or fexaramine treatment. (B) Bar graph ofserum alanine aminotransferase (ALT) levels in vehicle and fexaraminetreated mice after 3 weeks of an ethanol diet (C) Liver triglyceridelevels in vehicle and fexaramine treated mice after 3 weeks of anethanol or isocaloric (control) diet.

FIG. 45 is a graph of percentage activation of FXR versus the log valueof concentration for a retest of NSSK00024, fexaramine and DMSO.

FIG. 46 is a graph of percentage activation of FXR versus the log valueof concentration for a retest of NSSK00027, fexaramine and DMSO.

FIG. 47 is a graph of percentage activation of FXR versus the log valueof concentration for a retest of NSSK00089, fexaramine and DMSO.

FIG. 48 is a graph of percentage activation of FXR versus the log valueof concentration for a retest of NSSK00096, fexaramine and DMSO.

FIG. 49 is a graph of percentage activation of FXR versus the log valueof concentration for a retest of NSSK00110, fexaramine and DMSO.

FIG. 50 is a graph of percentage activation of FXR versus the log valueof concentration for a retest of deuterated feraramine, fexaramine andDMSO.

SEQUENCE LISTING

The amino acid sequences are shown using standard three letter code foramino acids, as defined in 37 C.F.R. 1.822.

SEQ ID NO. 1 is a protein sequence of GLP-1-(7-36).

SEQ ID NO. 2 is a protein sequence of GLP-2.

DETAILED DESCRIPTION I. TERMS

The following explanations of terms and methods are provided to betterdescribe the present disclosure and to guide those of ordinary skill inthe art in the practice of the present disclosure. The singular forms“a,” “an,” and “the” refer to one or more than one, unless the contextclearly dictates otherwise. For example, the term “comprising a FXRagonist” includes single or plural FXR agonists and is consideredequivalent to the phrase “comprising at least one FXR agonist.” The term“or” refers to a single element of stated alternative elements or acombination of two or more elements, unless the context clearlyindicates otherwise. As used herein, “comprises” means “includes.” Thus,“comprising A or B,” means “including A, B, or A and B,” withoutexcluding additional elements. Dates of GenBank® Accession Nos. referredto herein are the sequences available at least as early as March 13,2014. All references, including patents and patent applications, andGenBank® Accession numbers cited herein are incorporated by reference.

Unless explained otherwise, all technical and scientific terms usedherein have the same meaning as commonly understood to one of ordinaryskill in the art to which this disclosure belongs. Although methods andmaterials similar or equivalent to those described herein can be used inthe practice or testing of the present disclosure, suitable methods andmaterials are described below. The materials, methods, and examples areillustrative only and not intended to be limiting.

All groups herein are understood to include substituted groups unlessspecifically stated otherwise, or context indicates otherwise. Asubstituted group means that one or more hydrogen atoms of the specifiedgroup or radical is each, independently of one another, replaced withthe same or different non-hydrogen substituent. Exemplary substituentgroups are identified below.

Substituent groups for substituting for one or more hydrogens (any twohydrogens on a single carbon can be replaced with ═O, ═NR⁷⁰, ═N—OR⁷⁰,═N₂ or ═S) on saturated carbon atoms in the specified group or radicalare, unless otherwise specified, —R⁶⁰, halo, deuterium, ═O, —OR⁷⁰,—SR⁷⁰, —NR⁸⁰R⁸⁰, trihalomethyl, —CN, —OCN, —SCN, —NO, —NO₂, ═N₂, —N₃,—SO₂R⁷⁰, —SO₂O⁻ M⁺, —SO₂OR⁷⁰, —OSO₂R⁷⁰, —OSO₂O M⁺, —OSO₂OR⁷⁰,—P(O)(O)2(M⁺)₂, —P(O)(OR⁷⁰)O M⁺, —P(O)(OR⁷⁰)₂, —C(O)R⁷⁰, —C(S)R⁷⁰,—C(NR⁷⁰)R⁷⁰, —C(O)O⁻M⁺, —C(O)OR⁷⁰, —C(S)OR⁷⁰, —C(O)NR⁸⁰R⁸⁰,—C(NR⁷⁰)NR⁸⁰R⁸⁰, —OC(O)R⁷⁰, —OC(S)R⁷⁰, —OC(O)O⁻M⁺, —OC(O)OR⁷⁰,—OC(S)OR⁷⁰, —NR⁷⁰C(O)R⁷⁰, —NR⁷⁰C(S)R⁷⁰, —NR⁷⁰CO₂ ⁻M⁺, —NR⁷⁰CO₂ ⁻R⁷⁰,—NR⁷⁰C(S)OR⁷⁰, —NR⁷⁰C(O)NR⁸⁰R⁸⁰, —NR⁷⁰C(NR⁷⁰)R⁷⁰ or —NR⁷⁰C(NR⁷⁰)NR⁸⁰R⁸⁰,where R⁶⁰ is selected from alkyl, alkenyl, cycloalkyl, heteroalkyl,heterocycloalkylalkyl, cycloalkylalkyl, aryl, arylalkyl, heteroaryl orheteroarylalkyl, each of which maybe optionally further substituted;each R⁷⁰ is independently hydrogen or R⁶⁰; each R⁸⁰ is independently R⁷⁰or alternatively, two R⁸⁰'s, taken together with the nitrogen atom towhich they are bonded, form a 5-, 6- or 7-membered heterocycloalkylwhich may optionally include from 1 to 4 of the same or differentadditional heteroatoms selected from O, N or S, of which N may have —Hor C₁-C₃ alkyl substitution; and each M⁺ is a counter ion with a netsingle positive charge. Each M⁺ may independently be, for example, analkali ion, such as K⁺, Na⁺, Li⁺; an ammonium ion, such as ⁺N(R⁶⁰)₄; oran alkaline earth ion, such as [Ca²⁺]_(0.5), [Mg²⁺]_(0.5), or[Ba²⁺]_(0.5) (“subscript 0.5” means e.g. that one of the counter ionsfor such divalent alkali earth ions can be an ionized form of a compoundof the invention and the other a typical counter ion such as chloride,or two ionized compounds of the invention can serve as counter ions forsuch divalent alkali earth ions, or a doubly ionized compound of theinvention can serve as the counter ion for such divalent alkali earthions). As specific examples, —NR⁸⁰R⁸⁰ is meant to include —NH₂,—NH-alkyl, N-pyrrolidinyl, N-piperazinyl, 4N-methyl-piperazin-1-yl,N-morpholinyl and —N(alkyl)₂ such as, for example, —N(methyl)₂ or—N(methyl)(ethyl).

Substituent groups for hydrogens on unsaturated carbon atoms in“substituted” alkene, cycloalkene, alkyne, aryl and heteroaryl groupsare, unless otherwise specified, —R⁶⁰, halo, deuterium, —O⁻M⁺, —OR⁷⁰,—SR⁷⁰, —S⁻M⁺, —NR⁸⁰R⁸⁰, trihalomethyl, —CF₃, —CN, —OCN, —SCN, —NO, —NO₂,—N₃, —SO₂R⁷⁰, —SO₃ ⁻M⁺, —SO₃R⁷⁰, —OSO₂R⁷⁰, —OSO₃ ⁻M⁺, —OSO₃R⁷⁰, —PO₃⁻2(M⁺)₂, —P(O)(OR⁷⁰)O⁻M⁺, —P(O)(OR⁷⁰)₂, —C(O)R⁷⁰, —C(S)R⁷⁰, —C(NR⁷⁰)R⁷⁰,—CO₂ ⁻M⁺, —CO₂R⁷⁰—C(S)OR⁷⁰, —C(O)NR⁸⁰R⁸⁰, —C(NR⁷⁰)NR⁸⁰R⁸⁰, —OC(O)R⁷⁰,—OC(S)R⁷⁰, —OCO₂ ⁻M⁺, —OCO₂R⁷⁰, —OC(S)OR⁷⁰, —NR⁷⁰C(O)R⁷⁰, —NR⁷⁰C(S)R⁷⁰,—NR⁷⁰CO₂ ⁻M⁺, —NR⁷⁰CO₂R⁷⁰, —NR⁷⁰C(S)OR⁷⁰, —NR⁷⁰C(O)NR⁸⁰R⁸⁰,—NR⁷⁰C(NR⁷⁰)R⁷⁰ or —NR⁷⁰C(NR⁷⁰)NR⁸⁰R⁸⁰, where R⁶⁰, R⁷⁰, R⁸⁰ and M⁺ areas previously defined, provided that in case of substituted alkene oralkyne, the substituents are not —O⁻M⁺, —OR⁷⁰, —SR⁷⁰, or —S⁻M⁺.

Substituent groups for replacing hydrogens on nitrogen atoms in“substituted” heterocyclic groups are, unless otherwise specified, —R⁶⁰,—O⁻M⁺, —OR⁷⁰, —SR⁷⁰, —S⁻M⁺, —NR⁸⁰R⁸⁰, trihalomethyl, —CF₃, —CN, —NO,—NO₂, —S(O)₂R⁷⁰, —S(O)₂O⁻M⁺, —S(O)₂OR⁷⁰, —OS(O)₂R⁷⁰, —OS(O)₂O⁻M⁺,OS(O)₂OR⁷⁰, —P(O)(O⁻)₂(M⁺ ₂, —P(O(OR⁷⁰)O⁻M⁺, —P(O)(OR⁷⁰)(OR⁷⁰),—C(O)R⁷⁰, —C(S)R⁷⁰, —C(NR⁷⁰)R⁷⁰, —C(O)OR⁷⁰, —C(S)OR⁷⁰, —C(O)NR⁸⁰R⁸⁰,—C(NR⁷⁰)NR⁸⁰R⁸⁰, —OC(O)R⁷⁰, —OC(S)R⁷⁰, —OC(O)OR⁷⁰, —OC(S)OR⁷⁰,—NR⁷⁰C(O)R⁷⁰, —NR⁷⁰C(S)R⁷⁰, —NR⁷⁰C(O)OR⁷⁰, —NR⁷⁰C(S)OR⁷⁰,—NR⁷⁰C(O)NR⁸⁰R⁸⁰, —NR⁷⁰C(NR⁷⁰)R⁷⁰ or —NR⁷⁰C(NR⁷⁰)NR⁸⁰R⁸⁰, where R⁶⁰,R⁷⁰, R⁸⁰ and M⁺ are as previously defined.

In a preferred embodiment, a group that is substituted has 1substituent, 1 or 2 substituents, 1, 2, or 3 substituents or 1, 2, 3 or4 substituents.

Also, it is understood that the above definitions are not intended toinclude impermissible substitution patterns. Such impermissiblesubstitution patterns are understood by a person having ordinary skillin the art.

Additionally, it is understood by a person of ordinary skill in the artthat if an atom does not appear to have sufficient specific bonds tosatisfy valence requirements, such as an apparent trivalent carbon,there are sufficient implicit hydrogens present to satisfy those valencerequirements.

As used herein, the wavyline “

” indicates the point of attachment for a group or radical.

“Alkyl” refers to a hydrocarbon group having a saturated carbon chain,which, unless otherwise specified, may optionally be substituted,particularly with substituents as described in the definition of“substituted.” The chain may be cyclic, branched or unbranched. The term“lower alkyl” means that the alkyl chain includes 1-10 carbon atoms e.g.methyl, ethyl, n-propyl, isopropyl, n-butyl, iso-butyl, sec-butyl,tert-butyl, n-pentyl, tert-pentyl, neopentyl, isopentyl, sec-pentyl,3-pentyl, hexyl, heptyl, octyl, nonyl or decyl. Also, by way of example,a methyl group, an ethyl group, an n-propyl and an isopropyl group areall represented by the term C₁₋₃ alkyl. Likewise terms indicating largernumerical ranges of carbon atoms are representative of any linear orbranched hydrocarbyl falling within the numerical range. Thisinclusiveness applies to other hydrocarbyl terms bearing such numericalranges. The terms alkenyl and alkynyl refer to hydrocarbon groups havingcarbon chains containing one or more double or triple bonds,respectively.

“Alkylene” refers to divalent saturated aliphatic hydrocarbyl groupspreferably having from 1 to 10 carbon atoms, more preferably 1 to 4carbon atoms, that are either straight-chained or branched, which mayoptionally be substituted, particularly with substituents as describedherein, unless otherwise specified. This term is exemplified by groupssuch as methylene (—CH₂—), ethylene (—CH₂CH₂—), n-propylene(—CH₂CH₂CH₂), iso-propylene (—CH₂CH(CH₃)—) or (—CH(CH₃)CH₂—), and thelike.

“Alkenylene” refers to divalent unsaturated aliphatic hydrocarbyl groupspreferably having from 2 to 10 carbon atoms, more preferably 2 to 4carbon atoms, that are either straight-chained or branched, and includeat least one double bond. Unless otherwise specified, the group may beoptionally be substituted, particularly with substituents as describedherein. This term is exemplified by groups such as ethenylene (—CH═CH—)and propenylene (—CH═CHCH₂—) and the like.

“Alkynylene” refers to divalent unsaturated aliphatic hydrocarbyl groupspreferably having from 2 to 10 carbon atoms, more preferably 2 to 4carbon atoms, that are either straight-chained or branched and includeat least one triple bond. Unless otherwise specified, the group may beoptionally be substituted, particularly with substituents as describedherein. This term is exemplified by groups such as ethynylene (—C≡C—)and n-propynylene (—C≡CCH₂—) and the like.

“Alkylthio” refers to the group —S-alkyl.

“Alkoxy” refers to the group —O-alkyl. Alkoxy includes, by way ofexample, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, t-butoxy,sec-butoxy, n-pentoxy, and the like.

“Acyl” refers to the groups H—C(O)—, alkyl-C(O)—, alkenyl-C(O),alkynyl-C(O)—, cycloalkyl-C(O)—, cycloalkenyl-C(O)—, aryl-C(O)—,heteroaryl-C(O)—, or heterocyclic-C(O)—. By way of example, “acyl”includes the “acetyl” group CH₃C(O)—.

“Amino” refers to the group —NR²¹R²², wherein R²¹ and R²² independentlyare selected from hydrogen, alkyl, alkenyl, alkynyl, aryl, cycloalkyl,cycloalkenyl, heteroaryl, or heterocyclic, or where R²¹ and R²² areoptionally joined together with the nitrogen bound thereto to form aheterocyclic group.

“Aminocarbonyl” refers to the group —C(O)NR²¹R²², wherein R²¹ and R²²independently are selected from hydrogen, alkyl, alkenyl, alkynyl, aryl,cycloalkyl, cycloalkenyl, heteroaryl, or heterocyclic, or where R²¹ andR²² are optionally joined together with the nitrogen bound thereto toform a heterocyclic group.

“Aminosulfonyl” refers to the group —SO₂NR²¹R²² where R²¹ and R²² areindependently are selected from hydrogen, alkyl, alkenyl, alkynyl, aryl,cycloalkyl, cycloalkenyl, heteroaryl, or heterocyclic, or where R²¹ andR²² are optionally joined together with the nitrogen bound thereto toform a heterocyclic group.

“Aryl” or “Ar” refers to an aromatic moiety, such as a carbocyclic groupof from 6 to 15 carbon atoms having a single ring (e.g., phenyl) ormultiple condensed rings (e.g., naphthyl or anthryl) in which at leastone of the condensed rings is aromatic (e.g., 2-benzoxazolinone,2H-1,4-benzoxazin-3(4H)-one-7-yl, 9,10-dihydrophenanthrene, and thelike), provided that the point of attachment is through an atom of thearomatic aryl group. Unless otherwise specified, the aryl group may beoptionally be substituted, particularly with substituents as describedherein. Preferred aryl groups include phenyl and naphthyl.

“Alkenyl” refers to straight chain or branched hydrocarbyl groups havingfrom 2 to 6 carbon atoms and preferably 2 to 4 carbon atoms and havingat least 1 double bond. Unless otherwise specified, the alkenyl groupmay be optionally substituted. Such groups are exemplified, for example,bi-vinyl, allyl, and but-3-en-1-yl. Included within this term are thecis and trans isomers or mixtures of these isomers, unless otherwisespecified.

“Alkynyl” refers to straight chain or branched hydrocarbyl groups havingfrom 2 to 6 carbon atoms, and preferably 2 to 4 carbon atoms, and havingat least 1 site of triple bond unsaturation. Unless otherwise specified,the alkynyl group may be optionally substituted. Such groups areexemplified, for example, by ethynyl, 1-propynyl and 2-propynyl.

“Boronic acid” refers to the groups —B(OR)₂, where each R independentlyis selected from H, alkyl, cycloalkyl, aryl or where the R substituentsform a ring, such as in a picolinate ester

or a catechol ester

“Cycloalkyl” refers to a cyclic alkyl group of from 3 to 10 carbon atomshaving a single ring, which, unless otherwise specified, may beoptionally substituted. Examples of suitable cycloalkyl groups include,for instance, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,cycloheptyl, cyclooctyl and the like.

“Cycloalkenyl” refers to a cyclic alkenyl group of from 3 to 10 carbonatoms having a single ring, which, unless otherwise specified, may beoptionally substituted. Examples of suitable cycloalkenyl groupsinclude, for instance, cyclohexenyl, cyclopentenyl, and cyclobutenyl.

“Halo”, “halide” or “halogen” refers to fluoro, chloro, bromo, and iodoand is preferably fluoro or chloro.

“Hydroxy” or “hydroxyl” refers to the group —OH.

“Heteroaryl” refers to an aromatic group having from 1 to 10 carbonatoms and at least one, and more typically 1 to 4, heteroatoms selectedfrom oxygen, nitrogen or sulfur within the ring. Unless otherwisespecified, the heteroaryl group may be optionally substituted. Suchheteroaryl groups can have a single ring (e.g., pyridinyl, imidazolyl orfuryl) or multiple condensed rings (e.g., indolizinyl, quinolinyl,benzimidazolyl, benzopyrazolyl or benzothienyl), wherein at least one ofthe condensed rings is aromatic and may or may not contain a heteroatom,provided that the point of attachment is through an atom of an aromaticring. In one embodiment, the nitrogen and/or sulfur ring atom(s) of theheteroaryl group are optionally oxidized to provide N-oxide (N→O),sulfinyl, or sulfonyl moieties. Preferred heteroaryls include pyridinyl,pyrrolyl, indolyl, thiophenyl, benzopyrazolyl and furanyl.

“Heterocycle,” “heterocyclic,” “heterocycloalkyl,” and “heterocyclyl”refer to a saturated or unsaturated group having a single ring ormultiple condensed rings, including fused, bridged and spiro ringsystems, and having from 3 to 15 ring atoms, including at least one, andmore typically 1 to 4, hetero atoms. The hetero atoms are selected fromnitrogen, sulfur, or oxygen. Unless otherwise specified, the group maybe optionally substituted. In fused ring systems, one or more of therings can be cycloalkyl, aryl, or heteroaryl, provided that the point ofattachment is through a non-aromatic ring. In one embodiment, thenitrogen and/or sulfur atom(s) of the heterocyclic group are optionallyoxidized to provide for the N-oxide, —S(O)—, or —SO₂— moieties.

Examples of heterocycles and heteroaryls include, but are not limitedto, azetidine, pyrrole, imidazole, pyrazole, pyridine, pyrazine,pyrimidine, pyridazine, indolizine, isoindole, indole, dihydroindole,indazole, purine, quinolizine, isoquinoline, quinoline, phthalazine,naphthylpyridine, quinoxaline, quinazoline, cinnoline, pteridine,carbazole, carboline, phenanthridine, acridine, phenanthroline,isothiazole, phenazine, isoxazole, phenoxazine, phenothiazine,imidazolidine, imidazoline, piperidine, piperazine, indoline,phthalimide, 1,2,3,4-tetrahydroisoquinoline,4,5,6,7-tetrahydrobenzo[b]thiophene, thiazole, thiazolidine, thiophene,benzo[b]thiophene, morpholinyl, thiomorpholinyl (also referred to asthiamorpholinyl), 1,1-dioxothiomorpholinyl, piperidinyl, pyrrolidine,tetrahydrofuranyl, and the like.

“Nitro” refers to the group —NO₂.

“Polycyclic” refers to a saturated or unsaturated polycyclic ring systemhaving from about 5 to about 25 carbon atoms and having two or morerings (e.g. 2, 3, 4, or 5 rings). The rings can be fused and/or bridgedto form the polycyclic ring system, and unless otherwise specified, maybe optionally substituted. For example, the term includes bicyclo [4,5],[5,5], [5,6] or [6,6] ring systems, as well as the following bridgedring systems:

(i.e., [2.1.1], [2.2.1], [3.3.3], [4.3.1], [2.2.2], [4.2.2], [4.2.1],[4.3.2], [3.1.1], [3.2.1], [4.3.3], [3.3.2], [3.2.2], [3.3.1] and[4.1.1] polycyclic rings, respectively), and adamantyl. Polycyclicgroups can be linked to the remainder of the compound through anysynthetically feasible position. If a stereocenter is created then allpossible stereocenters are contemplated. Like the other polycarbocycles,these representative bicyclo and fused ring systems can optionallycomprise one or more double bonds in the ring system.

“Sulfonyl” refers to the group —SO₂—, and includes —SO₂-alkyl,—SO₂-alkenyl, —SO₂-cycloalkyl, —SO₂-cycloalkenyl, —SO₂aryl,—SO₂heteroaryl, or —SO₂-heterocyclic, wherein alkyl, alkenyl,cycloalkyl, cycloalkenyl, aryl, heteroaryl, and heterocyclic are asdefined herein. Sulfonyl includes groups such as methyl-SO₂,phenyl-SO₂—, and 4-methylphenyl-SO₂—.

The terms “carboxyl bioisosteric,” or “carboxyl bioisostere” refer to agroup with similar physical or chemical properties to a carboxyl groupthat produce broadly similar biological properties, but which may reducetoxicity or modify the activity of the compound, and may alter themetabolism of the compound. Exemplary carboxyl bioisosteres include, butare not limited to,

where X⁷, Y⁷, and Z⁷ are each independently selected from N, CH₂ or CO;

where X⁸ is selected from O, S or NMe;

where X⁹ is selected from O, N, S, CH or CH₂;

Additional carboxyl bioisosteric groups contemplated by the presentdisclosure include

Particular examples of the presently disclosed compounds include one ormore asymmetric centers; thus these compounds can exist in differentstereoisomeric forms. Accordingly, compounds and compositions may beprovided as individual pure enantiomers or as stereoisomeric mixtures,including racemic mixtures. In certain embodiments the compoundsdisclosed herein are synthesized in or are purified to be insubstantially enantiopure form, such as in a 90% enantiomeric excess, a95% enantiomeric excess, a 97% enantiomeric excess or even in greaterthan a 99% enantiomeric excess, such as in enantiopure form.

Prodrugs of the disclosed compounds also are contemplated herein. Aprodrug is an active or inactive compound that is modified chemicallythrough in vivo physiological action, such as hydrolysis, metabolism andthe like, into an active compound following administration of theprodrug to a subject. The term “prodrug” as used throughout this textmeans the pharmacologically acceptable derivatives such as esters,amides and phosphates, such that the resulting in vivo biotransformationproduct of the derivative is the active drug as defined in the compoundsdescribed herein. Prodrugs preferably have excellent aqueous solubility,increased bioavailability and are readily metabolized into the activeinhibitors in vivo. Prodrugs of a compounds described herein may beprepared by modifying functional groups present in the compound in sucha way that the modifications are cleaved, either by routine manipulationor in vivo, to the parent compound. The suitability and techniquesinvolved in making and using prodrugs are well known by those skilled inthe art. For a general discussion of prodrugs involving esters seeSvensson and Tunek, Drug Metabolism Reviews 165 (1988) and Bundgaard,Design of Prodrugs, Elsevier (1985).

The term “prodrug” also is intended to include any covalently bondedcarriers that release an active parent drug of the present invention invivo when the prodrug is administered to a subject. Since prodrugs oftenhave enhanced properties relative to the active agent pharmaceutical,such as, solubility and bioavailability, the compounds disclosed hereincan be delivered in prodrug form. Thus, also contemplated are prodrugsof the presently disclosed compounds, methods of delivering prodrugs andcompositions containing such prodrugs. Prodrugs of the disclosedcompounds typically are prepared by modifying one or more functionalgroups present in the compound in such a way that the modifications arecleaved, either in routine manipulation or in vivo, to yield the parentcompound. Prodrugs include compounds having a phosphonate and/or aminogroup functionalized with any group that is cleaved in vivo to yield thecorresponding amino and/or phosphonate group, respectively. Examples ofprodrugs include, without limitation, compounds having an acylated aminogroup, an ascorbate moiety, an ortho ester, an imidate group and/or aphosphonate ester or phosphonate amide group.

“Pharmaceutically acceptable salt” refers to pharmaceutically acceptablesalts of a compound, which salts are derived from a variety of organicand inorganic counter ions well known in the art and include, by way ofexample only, sodium, potassium, calcium, magnesium, ammonium,tetraalkylammonium, and the like. If the molecule contains a basicfunctionality, pharmaceutically acceptable salts include salts oforganic or inorganic acids, such as hydrochloride, hydrobromide,tartrate, mesylate, acetate, maleate, oxalate, and the like.

“Pharmaceutically acceptable excipient” refers to a substantiallyphysiologically inert substance that is used as an additive in apharmaceutical composition. As used herein, an excipient may beincorporated within particles of a pharmaceutical composition, or it maybe physically mixed with particles of a pharmaceutical composition. Anexcipient can be used, for example, as a carrier, flavoring, thickener,diluent, buffer, preservative, or surface active agent and/or to modifyproperties of a pharmaceutical composition. Examples of excipientsinclude, but are not limited, to polyvinylpyrrolidone (PVP), tocopherylpolyethylene glycol 1000 succinate (also known as vitamin E TPGS, orTPGS), dipalmitoyl phosphatidyl choline (DPPC), trehalose, sodiumbicarbonate, glycine, sodium citrate, and lactose.

“Enteric coating” refers to a coating such as may be applied todisclosed compounds or compositions comprising the compounds to helpprotect drugs from disintegration, digestion etc. in the stomach, suchas by enzymes or the pH of the stomach. Typically, the coating helpsprevent the drug from being digested in the stomach, and allows deliveryof the medication to the intestine.

The terms “administer,” “administering”, “administration,” and the like,as used herein, refer to methods that may be used to enable delivery ofagents or compositions to the desired site of biological action. Thesemethods include, but are not limited to oral routes, intraduodenalroutes and rectal administration. Administration techniques that areoptionally employed with the agents and methods described herein arefound in sources e.g., Goodman and Gilman, The Pharmacological Basis ofTherapeutics, current ed.; Pergamon; and Remington's, PharmaceuticalSciences (current edition), Mack Publishing Co., Easton, Pa. In certainembodiments, the agents and compositions described herein areadministered orally.

The term “calorie” refers to the amount of energy, e.g. heat, requiredto raise the temperature of 1 gram of water by 1° C. In various fieldssuch as medicine, nutrition, and the exercise sciences, the term“calorie” is often used to describe a kilocalorie. A kilocalorie is theamount of energy needed to increase the temperature of 1 kilogram ofwater by 1° C. One kilocalorie equals 1000 calories. The kilocalorie isabbreviated as kc, kcal or Cal, whereas the calorie or gram calorie isabbreviated as cal. In some embodiments, food intake in the subject ismeasured in terms of overall calorie consumption. Likewise, in someembodiments, fat intake can be measured in terms of calories from fat.

As used herein, the terms “co-administration,” “administered incombination with,” and their grammatical equivalents, are meant toencompass administration of the selected therapeutic agents to a singlepatient, and are intended to include treatment regimens in which theagents are administered by the same or different route of administrationor at the same or different times. In some embodiments the agentsdescribed herein will be co-administered with other agents. These termsencompass administration of two or more agents to the subject so thatboth agents and/or their metabolites are present in the subject at thesame time. They include simultaneous administration in separatecompositions, administration at different times in separatecompositions, and/or administration in a composition in which bothagents are present. Thus, in some embodiments, the agents describedherein and the other agent(s) are administered in a single composition.In some embodiments, the agents described herein and the other agent(s)are admixed in the composition.

The terms “effective amount,” “pharmaceutically effective amount” or“therapeutically effective amount” as used herein, refer to a sufficientamount of at least one agent being administered to achieve a desiredresult, e.g., to relieve to some extent one or more symptoms of adisease or condition being treated. In certain instances, the result isa reduction and/or alleviation of the signs, symptoms, or causes of adisease, or any other desired alteration of a biological system. Incertain instances, an “effective amount” for therapeutic uses is theamount of the composition comprising an agent as set forth hereinrequired to provide a clinically significant decrease in a disease. Anappropriate “effective” amount in any individual case can be determinedusing any suitable technique, such as a dose escalation study.

“Enhancing enteroendocrine peptide secretion” refers to a sufficientincrease in the level of the enteroendocrine peptide agent to, forexample, decrease hunger in a subject, to curb appetite in a subjectand/or decrease the food intake of a subject or individual and/or treatany disease or disorder described herein.

“FXR”: farnesoid X receptor (also known as nuclear receptor subfamily 1,group H, member 4 (NR1H4)) (OMIM: 603826): This protein functions as areceptor for bile acids, and when bound to bile acids, regulates theexpression of genes involved in bile acid synthesis and transport. FXRis expressed at high levels in the liver and intestine. Chenodeoxycholicacid and other bile acids are natural ligands for FXR. Similar to othernuclear receptors, when activated, FXR translocates to the cell nucleus,forms a dimer (in this case a heterodimer with RXR) and binds to hormoneresponse elements on DNA, which up- or down-regulates the expression ofcertain genes. One of the primary functions of FXR activation is thesuppression of cholesterol 7 alpha-hydroxylase (CYP7A1), therate-limiting enzyme in bile acid synthesis from cholesterol. FXR doesnot directly bind to the CYP7A1 promoter. Rather, FXR induces expressionof small heterodimer partner (SHP), which then functions to inhibittranscription of the CYP7A1 gene. In this way, a negative feedbackpathway is established in which synthesis of bile acids is inhibitedwhen cellular levels are already high. FXR sequences are publicallyavailable, for example from GenBank® sequence database (e.g., accessionnumbers NP_001193906 (human, protein) and NP_001156976 (mouse, protein),and NM_001206977 (human, nucleic acid) and NM_001163504 (mouse, nucleicacid)).

The term “metabolic disorder” refers to any disorder that involves analteration in the normal metabolism of carbohydrates, lipids, proteins,nucleic acids or a combination thereof. A metabolic disorder isassociated with either a deficiency or excess in a metabolic pathwayresulting in an imbalance in metabolism of nucleic acids, proteins,lipids, and/or carbohydrates. Factors affecting metabolism include, butare not limited to, the endocrine (hormonal) control system (e.g., theinsulin pathway, the enteroendocrine hormones including GLP-1, GLP-2,oxyntomodulin, PYY or the like), the neural control system (e.g., GLP-1in the brain) or the like. Examples of metabolic disorders include andare not limited to diabetes, insulin resistance, dyslipidemia, metabolicsyndrome, or the like.

The term “metabolic rate” refers to the rate at which the subject usesenergy. This is also known as the rate of metabolism, or the rate ofenergy consumption, and reflects the overall activity of theindividual's metabolism. The term basal metabolism refers to the minimumamount of energy required to maintain vital functions in an individualat complete rest, measured by the basal metabolic rate in a fastingindividual who is awake and resting in a comfortably warm environment.The term “basal metabolic rate” refers to the rate at which energy isused by an individual at rest. Basal metabolic rate is measured inhumans by the heat given off per unit time, and expressed as thecalories released per kilogram of body weight or per square meter ofbody surface per hour. The heart beating, breathing, maintaining bodytemperature, and other basic bodily functions all contribute to basalmetabolic rate. Basal metabolic rate can be determined to be the stablerate of energy metabolism measured in individuals under conditions ofminimum environmental and physiological stress, or essentially at restwith no temperature change. The basal metabolic rate among individualscan vary widely. One example of an average value for basal metabolicrate is about 1 calorie per hour per kilogram of body weight.

The terms “non-systemic” or “minimally absorbed” as used herein refer tolow systemic bioavailability and/or absorption of an administeredcompound. In some instances a non-systemic compound is a compound thatis substantially not absorbed systemically. In some embodiments, FXRagonist compositions described herein deliver an FXR agonist to thedistal ileum, colon, and/or rectum and not systemically (e.g., asubstantial portion of the FXR agonist administered is not systemicallyabsorbed). In some embodiments, the systemic absorption of anon-systemic compound is <0.1%, <0.3%, <0.5%, <0.6%, <0.7%, <0.8%,<0.9%, <1%, <1.5%, <2%, <3%, or <5% of the administered dose (wt. % ormol %). In some embodiments, the systemic absorption of a non-systemiccompound is <15% of the administered dose. In some embodiments, thesystemic absorption of a non-systemic compound is <25% of theadministered dose. In an alternative approach, a non-systemic FXRagonist is a compound that has lower systemic bioavailability relativeto the systemic bioavailability of a systemic FXR agonist. In someembodiments, the bioavailability of a non-systemic FXR agonist describedherein is <30%, <40%, <50%, <60%, or <70% of the bioavailability of asystemic FXR agonist. In some embodiments, the serum concentration ofthe FXR agonist in the subject remains below the compound's EC₅₀following administration.

The terms “prevent,” “preventing” or “prevention,” and other grammaticalequivalents as used herein, include preventing additional symptoms,preventing the underlying metabolic causes of symptoms, inhibiting thedisease or condition, e.g., arresting the development of the disease orcondition and are intended to include prophylaxis. The terms furtherinclude achieving a prophylactic benefit. For prophylactic benefit, thecompositions are optionally administered to a patient at risk ofdeveloping a particular disease, to a patient reporting one or more ofthe physiological symptoms of a disease, or to a patient at risk ofreoccurrence of the disease.

The term “subject”, “patient” or “individual” may be usedinterchangeably herein and refer to mammals and non-mammals, e.g.,suffering from a disorder described herein. Examples of mammals include,but are not limited to, any member of the mammalian class: humans,non-human primates such as chimpanzees, and other apes and monkeyspecies; farm animals such as cattle, horses, sheep, goats, swine;domestic animals such as rabbits, dogs, and cats; laboratory animalsincluding rodents, such as rats, mice and guinea pigs, and the like.Examples of non-mammals include, but are not limited to, birds, fish,amphibians, and the like. In one embodiment of the methods andcompositions provided herein, the mammal is a human.

The terms “treat,” “treating” or “treatment,” and other grammaticalequivalents as used herein, include alleviating, inhibiting or reducingsymptoms, reducing or inhibiting severity of, reducing incidence of,prophylactic treatment of, reducing or inhibiting recurrence of,preventing, delaying onset of, delaying recurrence of, abating orameliorating a disease or condition symptoms, ameliorating theunderlying metabolic causes of symptoms, inhibiting the disease orcondition, e.g., arresting the development of the disease or condition,relieving the disease or condition, causing regression of the disease orcondition, relieving a condition caused by the disease or condition, orstopping the symptoms of the disease or condition. The terms furtherinclude achieving a therapeutic benefit. Therapeutic benefit meanseradication or amelioration of the underlying disorder being treated,and/or the eradication or amelioration of one or more of thephysiological symptoms associated with the underlying disorder, suchthat an improvement is observed in the patient.

II. OVERVIEW

Disclosed herein are compounds that have activity as FXR agonists thatare structurally distinct from bile acids, other synthetic FXR ligands,and other natural FXR ligands. Also disclosed herein are embodiments ofa method for treating or preventing inflammation in the intestinesand/or a metabolic disorder, such as diabetes or obesity, byadministering a therapeutically effective amount of an FXR agonist tothe GI tract of a subject, such as one of the novel FXR agonistsdisclosed herein. Also disclosed herein are methods for treating orpreventing a cell proliferative disorder, such as cancer, for example inthe intestine, by administering a therapeutically effective amount of anFXR agonist to the subject (e.g., to the GI tract), such as one of thenovel FXR agonists disclosed herein. Also disclosed herein are methodsfor treating or preventing alcoholic liver disease (e.g., steatosis,cirrhosis, alcoholic hepatitis, elevated liver enzymes), such as in analcoholic subject, by administering a therapeutically effective amountof an FXR agonist to the subject (e.g., to the GI tract), such as one ofthe novel FXR agonists disclosed herein.

The absorption of these FXR agonists is substantially restricted to theintestinal lumen when delivered orally. In various embodiments,administration of one or more of the disclosed FXR agonists results inactivation of FXR transcriptional activity in the intestine, withoutsubstantially affecting other target tissues, such as liver or kidney.Surprisingly, despite this restricted activity, chronic administrationwith these agonists led to beneficial body-wide effects in obesesubjects. The disclosed FXR agonists have potent anti-obesity andglucose lowering effects in vivo. These effects have not been observedwith systemically-acting FXR ligands and include reductions in weightgain, hyperglycemia, and insulin resistance. In addition, administrationof these FXR agonists produced a beneficial, anti-inflammatory effect inthe intestines.

III. COMPOUNDS

Disclosed herein are embodiments of a compound having activity as an FXRagonist. Certain disclosed embodiments have formula 1

or a pharmaceutically acceptable salt thereof, wherein R is selectedfrom

is selected from aryl, heteroaryl, alkyl, alkenyl, cycloalkyl,heterocyclic, or polycyclic; R^(b) is selected from hydrogen, alkyl,alkenyl, or cycloalkyl; Y is CR^(g), N or N—O (N-oxide); R^(c), R^(d),R^(e) and R^(g) are each independently selected from hydrogen,deuterium, halide, alkyl, alkenyl, alkoxy, alkylthio, amino, sulfonyl,aminosulfonyl, aminocarbonyl, acyl, hydroxyl or nitro; R^(fa) and R^(fb)are each independently selected from hydrogen, deuterium, halide oralkyl; L^(a) and L^(b) are each independently selected from hydrogen,deuterium, alkyl or cycloalkyl, or together form a pi-bond; L^(c) andL^(d) are each independently selected from hydrogen, deuterium, alkyl orcycloalkyl; W is selected from O or —(C(L^(c))(L^(d)))_(s)-; s is 1, 2,3, 4, 5 or 6; n is 0 or 1; and X is aryl, heterocyclic or heteroaryl. Insome embodiments when R^(b) is hydrogen, the compounds have activity asFXR agonists. In other embodiments when R^(b) is hydrogen, the compoundsmay have reduced or substantially no activity as FXR agonists.

Also with reference to formula 1, the following provisos apply:

if W is CH₂ and L^(c) and L^(d) are both H, then X is not a benzopyran;

if R is

L^(c) and L^(d) are both H, and L^(a) and L^(b) are both H or togetherform a pi-bond, then X is not a benzopyran;

X is not substituted with —R^(x)-L^(x)-R^(x2), where R^(x) is selectedfrom O, NR^(x3), sulfonyl or S; R^(x3) is selected from H, alkyl,alkenyl, alkynyl, cycloalkyl, cycloalkenyl, or aryl; L^(x) is selectedfrom a bond, alkylene, alkenylene, alkynylene, cycloalkyl, cycloalkenyl,heterocyclic, aryl, heteroaryl or CR^(x4)R^(x5); R^(x4) and R^(x5) areeach independently selected from H, D, halogen, alkyl, alkenyl, alkynyl,cycloalkyl, cycloalkenyl, —C(O)OR^(x6), or —C(O)NR^(x6)R^(x7); R^(x6)_(and R) ^(x7) are each independently selected from H, alkyl, alkenyl,alkynyl, cycloalkyl or cycloalkenyl; R^(x2) is selected from—C(O)L^(x2)R^(x8) or a carboxyl bioisostere; L^(x2) is a bond orNR^(x3); R^(x8) is H, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl,—OR^(x9), N(R^(x9))₂, —C(O)R^(x9), —S(O)₂R^(x9), —C(O)OR^(x9),—S(O)₂N(R^(x9))₂ or C(O)N(R^(x9))₂; and each R^(x9) is independentlyselected from H, alkyl, alkenyl, alkynyl, cycloalkyl or cycloalkenyl;and

when R is

R^(c), R^(d), R^(e) and R^(fa) are all hydrogen, Y is CH and L^(a) andL^(b) are both H or together form a pi-bond, then

-   -   if R^(a) is cyclohexyl, R^(b) is methyl, and R^(fb) is H then X        is not phenyl, 4-biphenyl, 4-bromophenyl, 3-bromophenyl,        2-bromophenyl, 4-tert-butylphenyl, 3-methoxyphenyl,        3,5-dimethoxyphenyl, 3-(trifluoromethyl)phenyl,        4-(3,4-difluorophenyl)phenyl, 4-(3-acetylphenyl)phenyl,        4-(4-methylthiophenyl)phenyl, 4-(4-methoxyphenyl)phenyl,        4-(3-methoxyphenyl)phenyl, 4-(2-methoxyphenyl)phenyl,        4-(3,5-dichlorophenyl)phenyl, 4-(4-tert-butylphenyl)phenyl,        4-(3-ethoxyphenyl)phenyl, 4-(3-chlorophenyl)phenyl,        4-(3-methylphenyl)phenyl, 4-(4-methylphenyl)phenyl,        4-(2-methoxy-5-chlorophenyl)phenyl,        4-(3-chloro-4-fluorophenyl)phenyl,        4-(4-trifluoromethoxyphenyl)phenyl,        4-(3-trifluoromethoxyphenyl)phenyl,        4-(2,6-dimethoxyphenyl)phenyl, 4-(4-dimethylaminophenyl)phenyl,

-   -   if R^(a) is cyclohexyl, R^(fb) is H and X is

then R^(b) is not methyl, ethyl or tert-butyl;

-   -   if R^(b) is methyl, R^(fb) is H and X is

then R^(a) is not cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl;

-   -   if R^(a) is cyclohexyl, R^(fb) is H and X is

then R^(b) is not methyl or tert-butyl;

-   -   if R^(a) is cyclohexyl, R^(b) is methyl, R^(fb) is H and X is

then R^(h) is not hydroxyl, (trimethylsilyl)ethoxymethyl-O, methoxy,0-benzyl, OCH₂CO₂Et, OC(O)CH₃, OC(O)Ph or OSO₂CH₃;

-   -   if R^(a) is cyclohexyl, R^(b) is methyl, R^(fb) is H and X is

then R^(h) is not —CH═CHC(O)OMe, —CH═CHC(O)OEt, —CH═CHC(O)NMe₂,—CH═CHC(O)NH^(t)Bu, —CH═CHC(O)O^(t)Bu, —CH═CHC(O)O^(i)Pr,—CH═CHC(O)OCH₂Ph, —CH═CHC(O)OH, —CH═CHCH₂OMe, —CH═CHCH₂OEt or—CH═CHCH₂OPh.

In some embodiments of formula 1, R^(b) is substituted with substituentsthat improve the compounds water solubility. In certain embodiments,R^(b) is selected from alkyl, alkenyl, or cycloalkyl, each substitutedwith one or more hydroxyl groups.

In some embodiments, R^(a) is substituted with one or more hydroxylgroups, or a lower PEG group, such as PEG 2, PEG 3, PEG 4, PEG 5, PEG 6,PEG 8, PEG 10.

In some embodiments, X is not a benzopyran.

In particular embodiments, R is

leading to compounds having formula 2

In some disclosed embodiments, the compounds having activity as FXRagonists have general formula 3

or a pharmaceutically acceptable salt thereof. With reference to formula3, R¹ is selected from aryl, heteroaryl, heterocyclic, alkyl, alkenyl,cycloalkyl, cycloalkenyl or polycyclic; R² is selected from hydrogen,alkyl, alkenyl, or cycloalkyl; Y is selected from N, N—O (N-oxide) orC—R^(3d); R^(3a), R^(3b), R^(3c) and R^(3d) are each independentlyselected from hydrogen, deuterium, halide, alkyl, alkenyl, alkoxy,alkylthio, amino, sulfonyl, aminosulfonyl, aminocarbonyl, acyl, hydroxylor nitro; R^(4a) and R^(4b) are each independently selected fromhydrogen (H), deuterium (D), halide or alkyl; L¹ and L² areindependently selected from hydrogen, deuterium, alkyl, cycloalkyl, ortogether form a pi-bond; and R^(5a), R^(5b), R^(5c), R^(5d) and R^(5e)are each independently selected from hydrogen, deuterium, halide, alkyl,alkenyl, alkoxy, alkylthio, amino, sulfonyl, aminosulfonyl,aminocarbonyl, acyl, aryl, heteroaryl, cycloalkyl, heterocyclyl,hydroxyl or nitro, or any two adjacent groups selected together form anaryl, heteroaryl, cycloalkyl or heterocyclic ring. In some embodimentswhen R² is hydrogen, the compounds have activity as FXR agonists. Inother embodiments when R² is hydrogen, the compounds may have reduced orsubstantially no activity as FXR agonists.

Also with reference to formula II, none of R^(5a), R^(5b), R^(5c),R^(5d) or R^(5e) is —R^(x)-L^(x)-R^(x2), where R^(x) is selected from O,NR^(x3), sulfonyl or S; R^(x3) is selected from H, alkyl, alkenyl,alkynyl, cycloalkyl, cycloalkenyl, or aryl; L^(x) is selected from abond, alkylene, alkenylene, alkynylene, cycloalkyl, cycloalkenyl,heterocyclic, aryl, heteroaryl or CR^(x4)R^(x5); R^(x4) and R^(x5) areeach independently selected from H, D, halogen, alkyl, alkenyl, alkynyl,cycloalkyl, cycloalkenyl, —C(O)OR^(x6), or —C(O)NR^(x6)R^(x7); R^(x6)and R^(x7) are each independently selected from H, alkyl, alkenyl,alkynyl, cycloalkyl or cycloalkenyl; R^(x2) is selected from—C(O)L^(x2)R^(x8) or a carboxyl bioisostere; L^(x2) is a bond orNR^(x3); R^(x8) is H, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl,—OR^(x9), N(R^(x9))₂, —C(O)R^(x9), —S(O)₂R^(x9), —C(O)OR^(x9),—S(O)₂N(R^(x9))₂ or —C(O)N(R^(x9))₂; and each R^(x9) is independentlyselected from H, alkyl, alkenyl, alkynyl, cycloalkyl or cycloalkenyl;and

if L¹ and L² are both hydrogen or together form a pi-bond then at leastone of the following conditions applies: Y is N or C-halogen; or R¹ ispolycyclic; or R^(4a) is D; or R^(5a) is F, Cl, I; or R^(5d) and R^(5e)together form an aryl, heteroaryl, cycloalkyl or heterocyclic ring; orR^(5b) and R^(5c) together form an aryl, cycloalkyl, nitrogen-containingheterocyclic or nitrogen-containing heteroaryl ring, or any combinationthereof.

In some embodiments, R² is substituted with one or more groups thatimprove the compounds water solubility. In certain embodiments, R² issubstituted with one or more hydroxyl groups.

In some embodiments, one or more of R^(5a), R^(5b), R^(5c), R^(5d) orR^(5e) is selected from

where R^(5h) is alkyl, alkenyl, hydrogen, cycloalkyl, or heterocyclic.

In particular embodiments, L¹ and L² together form a pi-bond, leading tocompounds having a formula 4

In some embodiments of general formula 4, Y is CR^(3d), leading tocompounds having general formula 5

With reference to formula 5, R^(3d) or R^(5a) or both are halogen, suchas F, Cl, Br or I, and R¹, R², R^(3a), R^(3b), R^(3c), R^(4a), R^(4b),R^(5b), R^(5c), R^(5d) and R^(5e) are defined as for formula 3, above.In some working embodiments, R^(3d) or R^(5a) or both are F.

In other embodiments of formula 4, Y is N, resulting in compounds havinggeneral formula 6

where R¹, R², R^(3a), R^(3b), R^(3b), R^(3c), R^(4a), R^(4b), R^(5a),R^(5b), R^(5c), R^(5d) and R^(5e) are defined as for formula 3.

In certain embodiments of formula 4, R¹ is polycyclic. This leads tocompounds having general formula 7

With reference to formula 7, R², R^(3a), R^(3b), R^(3c), R^(4a), R^(4b),R^(5a), R^(5b), R^(5c), R^(5d), R^(5e) and Y are defined as for formula3, above. In some examples, the polycyclic is selected from

or adamantyl. In other examples, the polycyclic is selected from[2.1.1], [2.2.1], [3.3.3], [4.3.1], [2.2.2], [4.2.2], [4.2.1], [4.3.2],[3.1.1], [3.2.1], [4.3.3], [3.3.2], [3.2.2], [3.3.1], [4.1.1], oradamantyl. In certain working embodiments the polycyclic is

In certain embodiments of general formula 3, R^(5c) is anitrogen-containing heteroaryl ring. Exemplary nitrogen-containingheteroaryl rings include, but are not limited to, pyridine, pyrazole,pyrrole, imidazole, oxazole, isoxazole, thiazole, isothiazole, triazole,pyrimidine, pyrazine, triazine, benzopyrazole, benzimidazole, indole,quinoline, indazole, purine, quinoxaline, and acridine. In particularembodiments, the compounds have general formula 8

With reference to formula 8, R¹, R², R^(3a), R^(3b), R^(3c), R^(4a),R^(4b), R^(5a), R^(5b), R^(5e) and Y are defined as for formula 3,R^(6a), R^(6c), R^(6d) and R^(6g) are each independently selected fromH, D, halogen or alkyl, R^(6h) is selected from H, D, alkyl, cycloalkyl,aryl or heteroaryl, and Z is selected from N, CH or C-alkyl. In certainworking embodiments, Z is N and/or R^(6h) is methyl. In some examplesR^(6a), R^(6c), R^(6d) and R^(6g) are all hydrogen. In particularexamples, Y is C—R^(3d) and at least one of R^(3d) and R^(5a) is F.

In certain embodiments of general formula 5, R^(5c) is a 4-aminophenyl,leading to compounds having general formula 9

With reference to formula 9, R¹, R², R^(3a), R^(3b), R^(3c), R^(3d), R⁴,R^(5a), R^(5b), R^(5d) and R^(5e) are defined as for formula 5, R^(6a),R^(6b), R^(6c) and R^(6d) are each independently selected from H, D,halogen or alkyl, G is a lone pair of electrons or an oxygen, and R^(6e)and R^(6f) are each independently selected from alkyl, H or cycloalkyl,with the provisos that R^(3d) or R^(5a) or both are halogen, or R⁴ is D,or R¹ is polycyclic, or any combination thereof. In working embodiments,R^(6e) and R^(6f) are both methyl.

In certain embodiments, compounds having formula 9 are N-oxides, leadingto compounds having formula 10

In particular examples of any of the above embodiments R^(4a) is D,R^(4b) is H, and/or R² is methyl. In other examples, both R^(4a) andR^(4b) are D. And in particular embodiments of formulas 3, 4, 5, 6, 7, 9or 10, R¹ is cyclohexyl.

In some disclosed embodiments, the compounds having activity as FXRagonists have general formula 11

or a pharmaceutically acceptable salt thereof, wherein R^(a) is selectedfrom aryl, heteroaryl, alkyl, alkenyl, cycloalkyl, heterocyclic, orpolycyclic; R^(b) is selected from alkyl, alkenyl, or cycloalkyl; Y isCR^(g), N or N—O (N-oxide); R^(c), R^(d), R^(e) and R^(g) are eachindependently selected from hydrogen, deuterium, halide, alkyl, alkenyl,alkoxy, alkylthio, amino, sulfonyl, aminosulfonyl, aminocarbonyl,cycloalkyl, heterocyclic, acyl, hydroxyl or nitro; R^(fa) and R^(fb) areeach independently selected from hydrogen, deuterium, halide or alkyl; Xis aryl, heterocyclic or heteroaryl; R is selected from

L^(a) and L^(b) are each independently selected from hydrogen,deuterium, alkyl or cycloalkyl, or together form a pi-bond; L^(c) andL^(d) are each independently selected from hydrogen, deuterium, alkyl orcycloalkyl; W is selected from O or —(C(L^(c))(L^(d))_(s)-; s is 1, 2,3, 4, 5 or 6; n is 0 or 1; and X is not substituted with—R^(x)-L^(x)-R^(x2), where R^(x) is selected from O, NR^(x3), sulfonylor S; R^(x3) is selected from H, alkyl, alkenyl, alkynyl, cycloalkyl,cycloalkenyl, or aryl; L^(x) is selected from a bond, alkylene,alkenylene, alkynylene, cycloalkyl, cycloalkenyl, heterocyclic, aryl,heteroaryl or CR^(x4)R^(x5); R^(x4) and R^(x5) are each independentlyselected from H, D, halogen, alkyl, alkenyl, alkynyl, cycloalkyl,cycloalkenyl, —C(O)OR^(x6), or —C(O)NR^(x6)R^(x7); R^(x6) and R^(x7) areeach independently selected from H, alkyl, alkenyl, alkynyl, cycloalkylor cycloalkenyl; R^(x2) is selected from —C(O)L^(x2)R^(x8) or a carboxylbioisostere; L^(x2) is a bond or NR^(x3); R^(x8) is H, alkyl, alkenyl,alkynyl, cycloalkyl, cycloalkenyl, —OR^(x9), N(R^(x9))₂, —C(O)R^(x9),—S(O)₂R^(x9), —C(O)OR^(x9), —S(O)₂N(R^(x9))₂ or —C(O)N(R^(x9))₂; andeach R^(x9) is independently selected from H, alkyl, alkenyl, alkynyl,cycloalkyl or cycloalkenyl.

In some disclosed embodiments, the compounds having activity as FXRagonists have general formula 12

or a pharmaceutically acceptable salt thereof. With reference to formula12, R is selected from

L^(a) and L^(b) are each independently selected from hydrogen,deuterium, alkyl or cycloalkyl, or together form a pi-bond; L^(c) andL^(d) are each independently selected from hydrogen, deuterium, alkyl orcycloalkyl; W is selected from O or —(C(L^(c))(L^(d)))_(s)-; s is 1, 2,3, 4, 5 or 6; n is 0 or 1; R¹ is selected from aryl, heteroaryl,heterocyclic, alkyl, alkenyl, cycloalkyl, cycloalkenyl or polycyclic; R²is selected from alkyl, alkenyl, or cycloalkyl; Y is selected from N,N—O (N-oxide) or C—R^(3d); R^(3a), R^(3b), R^(3c) and R^(3d) are eachindependently selected from hydrogen, deuterium, halide, alkyl, alkenyl,alkoxy, alkylthio, amino, sulfonyl, aminosulfonyl, aminocarbonyl, acyl,hydroxyl or nitro; R^(4a) and R^(4b) are each independently selectedfrom hydrogen, deuterium, halide or alkyl; R^(5a), R^(5b), R^(5c),R^(5d) and R^(5e) are each independently selected from hydrogen,deuterium, halide, alkyl, alkenyl, alkoxy, alkylthio, amino, sulfonyl,aminosulfonyl, aminocarbonyl, acyl, aryl, heteroaryl, cycloalkyl,heterocyclyl, hydroxyl or nitro, or any two adjacent groups selectedtogether form an aryl, heteroaryl, cycloalkyl or heterocyclic ring; andnone of R^(5a), R^(5b), R^(5c), R^(5d) or R^(5e) is —R^(x)-L^(x)-R^(2x),where R^(x) is selected from O, NR^(x3), sulfonyl or S; R^(x3) isselected from H, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, oraryl; L^(x) is selected from a bond, alkylene, alkenylene, alkynylene,cycloalkyl, cycloalkenyl, heterocyclic, aryl, heteroaryl orCR^(x4)R^(x5); R^(x4) and R^(x5) are each independently selected from H,D, halogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl,—C(O)OR^(x6), or C(O)NR^(x6)R^(x7);R^(x6) and R^(x7) are eachindependently selected from H, alkyl, alkenyl, alkynyl, cycloalkyl orcycloalkenyl; R^(x2) is selected from —C(O)L^(x2)R^(x8) or a carboxylbioisostere; L^(x2) is a bond or NR^(x3); R^(x8) is H, alkyl, alkenyl,alkynyl, cycloalkyl, cycloalkenyl, —OR^(x9), N(R^(x9))₂, —C(O)R^(x9),—S(O)₂R^(x9), —C(O)OR^(x9), —S(O)₂N(R^(x9))₂ or —C(O)N(R^(x9))₂; andeach R^(x9) is independently selected from H, alkyl, alkenyl, alkynyl,cycloalkyl or cycloalkenyl.

In some other embodiments, the compounds having activity as FXR agonistshave general formula 13

or a pharmaceutically acceptable salt thereof, wherein R is selectedfrom

and L^(b) are each independently selected from hydrogen, deuterium,alkyl or cycloalkyl, or together form a pi-bond; L^(c) and L^(d) areeach independently selected from hydrogen, deuterium, alkyl orcycloalkyl; W is selected from O or —(C(L^(c))(L^(d)))_(s)-; s is 1, 2,3, 4, 5 or 6; n is 0 or 1; R^(b) is selected from alkyl, alkenyl, orcycloalkyl; Y is CR^(g), N or N—O (N-oxide); R^(c), R^(e) and R^(g) areeach independently selected from hydrogen, deuterium, halide, alkyl,alkenyl, alkoxy, alkylthio, amino, sulfonyl, aminosulfonyl,aminocarbonyl, cycloalkyl, heterocyclic, acyl, hydroxyl or nitro; R^(fa)and R^(fb) are each independently selected from hydrogen, deuterium,halide or alkyl; R^(h) and R^(j) are each independently selected fromhydrogen, deuterium, halide, alkyl, cycloalkyl, heterocycloalkyl,alkenyl, aryl or heteroaryl; X is aryl, heterocyclic or heteroaryl; andX is not substituted with —R^(x)-L^(x)-R^(x2), where R^(x) is selectedfrom O, NR^(x3), sulfonyl or S; R^(x3) is selected from H, alkyl,alkenyl, alkynyl, cycloalkyl, cycloalkenyl, or aryl; L^(x) is selectedfrom a bond, alkylene, alkenylene, alkynylene, cycloalkyl, cycloalkenyl,heterocyclic, aryl, heteroaryl or CR^(x4)R^(x5); R^(x4) and R^(x5) areeach independently selected from H, D, halogen, alkyl, alkenyl, alkynyl,cycloalkyl, cycloalkenyl, —C(O)OR^(x6), or C(O)NR^(x6)R^(x7); R^(x6) andR^(x7) are each independently selected from H, alkyl, alkenyl, alkynyl,cycloalkyl or cycloalkenyl; R^(x2) is selected from —C(O)L^(x2)R^(x8) ora carboxyl bioisostere; L^(x2) is a bond or NR^(x3); R^(x8) is H, alkyl,alkenyl, alkynyl, cycloalkyl, cycloalkenyl, —OR^(x9), N(R^(x9))₂,—C(O)R^(x9), —S(O)₂R^(x9), —C(O)OR^(x9), —S(O)₂N(R^(x9))₂ or—C(O)N(R^(x9))₂; and each R^(x9) is independently selected from H,alkyl, alkenyl, alkynyl, cycloalkyl or cycloalkenyl.

In some embodiments, prodrugs of compounds having activity as FXRagonists have general formula 14

or a pharmaceutically acceptable salt thereof, wherein R^(a) is selectedfrom aryl, heteroaryl, alkyl, alkenyl, cycloalkyl, heterocyclic, orpolycyclic; Rh is selected from alkyl, alkenyl, or cycloalkyl; Y isCR^(g), N or N—O (N-oxide); R^(c), R^(d), R^(e) and R^(g) are eachindependently selected from hydrogen, deuterium, halide, alkyl, alkenyl,alkoxy, alkylthio, amino, sulfonyl, aminosulfonyl, aminocarbonyl, acyl,hydroxyl or nitro; R^(fa) and R^(fb) are each independently selectedfrom hydrogen, deuterium, halide or alkyl; X is aryl, heterocyclic orheteroaryl; R^(y) and R^(z) are selected from alkyl, cycloalkyl,heterocyclic alkyl, aryl, or heteroaryl, or R^(y) and R^(z) may togetherform a cycloheteroalkyl ring; L^(a) and L^(b) are independently H, D oralkyl or together form a π-bond, a cyclopropyl or an epoxide ring; L^(c)and L^(d) are independently H, D or alkyl; and X is not substituted with—R^(x)-L^(x)-R^(x2), where R^(x) is selected from O, NR^(x3), sulfonylor S; R^(x3) is selected from H, alkyl, alkenyl, alkynyl, cycloalkyl,cycloalkenyl, or aryl; L^(x) is selected from a bond, alkylene,alkenylene, alkynylene, cycloalkyl, cycloalkenyl, heterocyclic, aryl,heteroaryl or CR^(x4)R^(x5); R^(x4) and R^(x5) are each independentlyselected from H, D, halogen, alkyl, alkenyl, alkynyl, cycloalkyl,cycloalkenyl, —C(O)OR^(x6), —C(O)NR^(x6)R^(x7); R^(x6) and R^(x7) areeach independently selected from H, alkyl, alkenyl, alkynyl, cycloalkylor cycloalkenyl; R^(x2) is selected from —C(O)L^(x2)R^(x8) or a carboxylbioisostere; L^(x2) is a bond or NR^(x3); R^(x8) is H, alkyl, alkenyl,alkynyl, cycloalkyl, cycloalkenyl, —OR^(x9), N(R^(x9))₂, —C(O)R^(x9),—S(O)₂R^(x9), —C(O)OR^(x9), —S(O)₂N(R^(x9))₂ or C(O)N(R^(x9))₂; and eachR^(x9) is independently selected from H, alkyl, alkenyl, alkynyl,cycloalkyl or cycloalkenyl.

In particular embodiments of formula 14, R^(y) and R^(z) together form a5-membered heteroalkyl ring substituted with an ascorbate moiety,leading to compounds having formula 15

In other embodiments, prodrugs of compounds having activity as FXRagonists have general formula 16

or a pharmaceutically acceptable salt thereof, wherein R^(a) is selectedfrom aryl, heteroaryl, alkyl, alkenyl, cycloalkyl, heterocyclic, orpolycyclic; R^(b) is selected from alkyl, alkenyl, or cycloalkyl; Y isCR^(g), N or N—O (N-oxide); R^(c), R^(d), R^(e) and R^(g) are eachindependently selected from hydrogen, deuterium, halide, alkyl, alkenyl,alkoxy, alkylthio, amino, sulfonyl, aminosulfonyl, aminocarbonyl, acyl,hydroxyl or nitro; R^(fa) and R^(fb) are each independently selectedfrom hydrogen, deuterium, halide or alkyl; X is aryl, heterocyclic orheteroaryl, L^(a) and L^(b) are independently H, D or alkyl or togetherform a π-bond, a cyclopropyl or an epoxide ring; L^(c) and L^(d) areindependently H, D or alkyl; and X is not substituted with—R^(x)-L^(x)-R^(x2), where R^(x) is selected from O, NR^(x3), sulfonylor S; R^(x3) is selected from H, alkyl, alkenyl, alkynyl, cycloalkyl,cycloalkenyl, or aryl; L^(x) is selected from a bond, alkylene,alkenylene, alkynylene, cycloalkyl, cycloalkenyl, heterocyclic, aryl,heteroaryl or CR^(x4)R^(x5); R^(x4) and R^(x5) are each independentlyselected from H, D, halogen, alkyl, alkenyl, alkynyl, cycloalkyl,cycloalkenyl, —C(O)OR^(x6), or —C(O)NR^(x6)R^(x7); R^(x6) and R^(x7) areeach independently selected from H, alkyl, alkenyl, alkynyl, cycloalkylor cycloalkenyl; R^(x2) is selected from —C(O)L^(x2)R^(x8) or a carboxylbioisostere; L^(x2) is a bond or NR^(x3); R^(x8) is H, alkyl, alkenyl,alkynyl, cycloalkyl, cycloalkenyl, —OR^(x9), N(R^(x9))₂, —C(O)R^(x9),—S(O)₂R^(x9), —C(O)OR^(x9), —S(O)₂N(R^(x9))₂ or —C(O)N(R^(x9))₂; andeach R^(x9) is independently selected from H, alkyl, alkenyl, alkynyl,cycloalkyl or cycloalkenyl.

In still further embodiments, prodrugs of compounds having activity asFXR agonists have general formula 17

or a pharmaceutically acceptable salt thereof, wherein R is selectedfrom

L^(a) and L^(b) are each independently selected from hydrogen,deuterium, alkyl or cycloalkyl, or together form a pi-bond; L^(c) andL^(d) are each independently selected from hydrogen, deuterium, alkyl orcycloalkyl; W is selected from O or —(C(L^(c))(L^(d)))_(s)-; s is 1, 2,3, 4, 5 or 6; n is 0 or 1; R^(a) is selected from aryl, heteroaryl,alkyl, alkenyl, cycloalkyl, heterocyclic, or polycyclic; R^(b) isselected from alkyl, alkenyl, or cycloalkyl; Y is CR^(g), N or N—O(N-oxide); R^(c), R^(d), R^(e) and R^(g) are each independently selectedfrom hydrogen, deuterium, halide, alkyl, alkenyl, alkoxy, alkylthio,amino, sulfonyl, aminosulfonyl, aminocarbonyl, cycloalkyl, heterocyclic,acyl, hydroxyl or nitro; R^(fa) and R^(fb) are each independentlyselected from hydrogen, deuterium, halide or alkyl; R^(k) and R^(m) areindependently selected from H, alkyl, aryl, cycloalkyl,heterocycloalkyl, heteroaryl, or together R^(k) and R^(m) form acycloalkyl or heterocycloalkyl ring; X is aryl, heterocyclic orheteroaryl; and X is not substituted with —R^(x)-L^(x)-R^(x2), whereR^(x) is selected from O, NR^(x3), sulfonyl or S; R^(x3) is selectedfrom H, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, or aryl;L^(x) is selected from a bond, alkylene, alkenylene, alkynylene,cycloalkyl, cycloalkenyl, heterocyclic, aryl, heteroaryl orCR^(x4)R^(x5); R^(x4) and R^(x5) are each independently selected from H,D, halogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl,—C(O)OR^(x6), or —C(O)NR^(x6)R^(x7); R^(x6) _(and R) ^(x7) are eachindependently selected from H, alkyl, alkenyl, alkynyl, cycloalkyl orcycloalkenyl; R^(x2) is selected from —C(O)L^(x2)R^(x8) or a carboxylbioisostere; L^(x2) is a bond or NR^(x3); R^(x8) is H, alkyl, alkenyl,alkynyl, cycloalkyl, cycloalkenyl, —OR^(x9), N(R^(x9))₂, —C(O)R^(x9),—S(O)₂R^(x9), —C(O)OR^(x9), —S(O)₂N(R^(x9))₂ or —C(O)N(R^(x9))₂; andeach R^(x9) is independently selected from H, alkyl, alkenyl, alkynyl,cycloalkyl or cycloalkenyl.

In some embodiments of formula 17 R^(k) and R^(m) together form a5-membered ring, leading to compounds having a formula 18

or pharmaceutically acceptable salt thereof, wherein each R^(n) isindependently selected from H, alkyl, or a metal salt such as Na, K, orLi.

In any of the above embodiments of formulas 1, 9 or 13-18, X isheteroaryl or heterocyclic, and in particular embodiments, X is pyridineor piperidine.

In other embodiments of formulas 1, 10, 11, or 13-18, X is a phenylsubstituted with an aryl or heteroaryl group. In certain embodiments, Xis a phenyl substituted with the aryl or heteroaryl group selected frombenzoxazine, dihydrobenzoxazine, quinoxaline, tetrahydroquinoxaline,benzodioxane, benzothiazine, dihydrobenzothiazine,dihydrobenzothiazine-1,1-dioxide, benzodithiine,benzodithiine-1,1,4,4-tetraoxide, benzofuran, benzothiophene, indole,benzisoxazole, indazole, benzotriazole, benzimidazole, benzoxazole,benzthiazole or benzisothiazole. In particular embodiments, X isselected from

A person of ordinary skill in the art will appreciate that compounds ofany of the above embodiments may have one or more stereocenter, and thateach stereocenter independently may have an R or S configuration.

A person of ordinary skill in the art will appreciate that prodrugcompounds satisfying one of formulas 14-18 may also have intrinsicactivity as FXR agonists, as well as acting as a prodrug for a compoundhaving FXR activity.

Exemplary working embodiments of compounds having activity as FXRagonists and satisfying one or more of the general formulas 1-18 areprovided below.

Other exemplary working embodiments include:

-   methyl    (E)-3-(3-(N-(4-(1-methyl-1H-indazol-5-yl)benzyl)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (E)-3-(3-((1R,2S,4S)—N-(4-(1-methyl-1H-indazol-5-yl)benzyl)bicyclo[2.2.1]heptane-2-carboxamido)phenyl)acrylate,-   methyl    (E)-3-(3-(1-methyl-N-(4-(1-methyl-1H-indazol-5-yl)benzyl)piperidine-4-carboxamido)phenyl)acrylate,-   methyl    (E)-3-(5-(N-((1-methyl-1H-benzo[f]indazol-8-yl)methyl)cyclohexanecarboxamido)pyridin-3-yl)acrylate,-   methyl    (E)-3-(3-fluoro-5-((1S,2R,4R)—N-((1-methyl-1H-benzo[f]indazol-8-yl)methyl)bicyclo[2.2.1]heptane-2-carboxamido)phenyl)acrylate,-   methyl    (E)-3-(3-(N-((9-fluoro-1-methyl-1H-benzo[f]indazol-8-yl)methyl)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (E)-3-(3-((1R,4S)—N-((9-fluoro-1-methyl-1H-benzo[f]indazol-8-yl)methyl)bicyclo[2.2.1]heptane-2-carboxamido)phenyl)acrylate,-   methyl    (E)-3-(3-(N-((9-fluoro-1-methyl-1H-benzo[f]indazol-8-yl)methyl)-1-methylpiperidine-4-carboxamido)phenyl)acrylate,-   methyl    (E)-3-(5-(N-((9-fluoro-1-methyl-1H-benzo[f]indazol-8-yl)methyl)cyclohexanecarboxamido)pyridin-3-yl)acrylate,-   methyl    (E)-3-(3-fluoro-5-(N-((9-fluoro-1-methyl-1H-benzo[f]indazol-8-yl)methyl)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (E)-3-(3-fluoro-5-((1R,4S)—N-((9-fluoro-1-methyl-1H-benzo[f]indazol-8-yl)methyl)bicyclo[2.2.1]heptane-2-carboxamido)phenyl)acrylate,-   methyl    (E)-3-(5-(N-((9-chloro-1-methyl-1H-benzo[f]indazol-8-yl)methyl)cyclohexanecarboxamido)pyridin-3-yl)acrylate,-   methyl    (E)-3-(5-((1R,4S)—N-((9-chloro-1-methyl-1H-benzo[f]indazol-8-yl)methyl)bicyclo[2.2.1]heptane-2-carboxamido)pyridin-3-yl)acrylate,-   methyl    (E)-3-(3-(N-((9-chloro-1-methyl-1H-benzo[f]indazol-8-yl)methyl)-1-methylpiperidine-4-carboxamido)-5-fluorophenyl)acrylate,-   methyl    (E)-3-(3-(N-((7-(dimethylamino)naphthalen-1-yl)methyl)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (E)-3-(3-(N-((7-(dimethylamino)-8-fluoronaphthalen-1-yl)methyl)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (E)-3-(3-(N-((4-(1-methyl-1H-indazol-5-yl)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (E)-3-(5-(N-((4-(1-methyl-1H-indazol-5-yl)phenyl)methyl-d)cyclohexanecarboxamido)pyridin-3-yl)acrylate,-   methyl    (E)-3-(5-(N-((2-fluoro-4-(1-methyl-1H-indazol-5-yl)phenyl)methyl-d)cyclohexanecarboxamido)pyridin-3-yl)acrylate,-   methyl    (E)-3-(3-fluoro-5-(N-((2-fluoro-4-(1-methyl-1H-indazol-5-yl)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (E)-3-(3-(N-((2-chloro-4-(1-methyl-1H-indazol-5-yl)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (E)-3-(3-((1S,2R,4R)—N-((2-chloro-4-(1-methyl-1H-indazol-5-yl)phenyl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)phenyl)acrylate,-   methyl    (E)-3-(3-(N-((2-chloro-4-(1-methyl-1H-indazol-5-yl)phenyl)methyl-d)cyclohexanecarboxamido)-5-fluorophenyl)acrylate,-   methyl    (E)-3-(5-((1S,2R,4R)—N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)pyridin-3-yl)acrylate,-   methyl    (E)-3-(3-(N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)cyclohexanecarboxamido)-5-fluorophenyl)acrylate,-   methyl    (E)-3-(3-((1S,2R,4R)—N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)-5-fluorophenyl)acrylate,-   methyl    (E)-3-(3-(N4-((4′-(dimethylamino)-3-fluoro-[1,1′-biphenyl]-4-yl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (E)-3-(5-(N-((4′-(dimethylamino)-3-fluoro-[1,1′-biphenyl]-4-yl)methyl-d)cyclohexanecarboxamido)pyridin-3-yl)acrylate,-   methyl    (E)-3-(5-((1S,2R,4R)—N-((4′-(dimethylamino)-3-fluoro-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)pyridin-3-yl)acrylate,-   methyl    (E)-3-(3-(N4-((3-chloro-4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (E)-3-(5-(N4(3-chloro-4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)cyclohexanecarboxamido)pyridin-3-yl)acrylate,-   methyl    (E)-3-(3-fluoro-5-(N-((2-fluoro-4-(1-methyl-1H-indazol-5-yl)phenyl)methyl-d)benzamido)phenyl)acrylate,-   methyl    (E)-3-(3-(N-((2-chloro-4-(1-methyl-1H-indazol-5-yl)phenyl)methyl-d)benzamido)-5-fluorophenyl)acrylate,-   methyl    (E)-3-(3-(N-((2-fluoro-4-(1-methyl-1H-indazol-5-yl)phenyl)methyl-d)benzamido)phenyl)acrylate,-   methyl    (E)-3-(5-((1S,2R,4R)—N-((2-fluoro-4-(1-methyl-1H-indazol-5-yl)phenyl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)pyridin-3-yl)acrylate,-   methyl    (E)-3-(3-(N-(2-chloro-4-(1-methyl-1H-indazol-5-yl)benzyl)-1-methylpiperidine-4-carboxamido)phenyl)acrylate,-   methyl    (E)-3-(3-((1S,2R,4R)—N-(2-chloro-4-(1-methyl-1H-indazol-5-yl)benzyl)bicyclo[2.2.    1]heptane-2-carboxamido)phenyl)acrylate,-   methyl    (E)-3-(3-((1S,2R,4R)—N-(2-chloro-4-(1-methyl-1H-indazol-5-yl)benzyl)bicyclo[2.2.    1]heptane-2-carboxamido)-5-fluorophenyl)acrylate,-   methyl    (E)-3-(5-((1S,2R,4R)—N-((2-chloro-4-(1-methyl-1H-indazol-5-yl)phenyl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)pyridin-3-yl)acrylate,-   methyl    (E)-3-(3-(N-(4-(1-methyl-1H-indazol-5-yl)benzyl)benzamido)phenyl)acrylate,-   methyl    (E)-3-(3-fluoro-5-(N-(4-(1-methyl-1H-indazol-5-yl)benzyl)benzamido)phenyl)acrylate,-   methyl    (E)-3-(3-(N-(2-fluoro-4-(1-methyl-1H-indazol-5-yl)benzyl)benzamido)phenyl)acrylate,-   methyl    (E)-3-(5-(N-(2-fluoro-4-(1-methyl-1H-indazol-5-yl)benzyl)benzamido)pyridin-3-yl)acrylate,-   methyl    (E)-3-(3-(N-((4-(1-methyl-1H-indazol-5-yl)phenyl)methyl-d)benzamido)phenyl)acrylate,-   methyl    (E)-3-(3-fluoro-5-(N-((4-(1-methyl-1H-indazol-5-yl)phenyl)methyl-d)benzamido)phenyl)acrylate,-   methyl    (E)-3-(5-(N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)cyclohexanecarboxamido)pyridin-3-yl)acrylate,-   methyl    (E)-3-(3-((1S,2R,4R)—N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl)bicyclo[2.2.1]heptane-2-carboxamido)phenyl)acrylate,-   methyl    (E)-3-(5-((1S,2R,4R)—N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl)bicyclo[2.2.1]heptane-2-carboxamido)pyridin-3-yl)acrylate,-   methyl    (E)-3-(3-(N-((3-chloro-4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)cyclohexanecarboxamido)-5-fluorophenyl)acrylate,-   methyl    (E)-3-(3-((1S,2R,4R)—N-((4′-(dimethylamino)-3-fluoro-[1,1′-biphenyl]-4-yl)methyl)bicyclo[2.2.1]heptane-2-carboxamido)phenyl)acrylate,-   methyl    (E)-3-(3-((1S,2R,4R)—N-((4′-(dimethylamino)-3-fluoro-[1,1′-biphenyl]-4-yl)methyl)bicyclo[2.2.1]heptane-2-carboxamido)-5-fluorophenyl)acrylate,-   methyl (E)-3-(3-((1S,2R,4R)—N-((3-chloro-4′-(dimethylamino)-[1,    1′-biphenyl]-4-yl)methyl)bicyclo[2.2.1]heptane-2-carboxamido)phenyl)acrylate,-   methyl    (E)-3-(3-((1S,2R,4R)—N-((3-chloro-4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)phenyl)acrylate,-   methyl    (E)-3-(5-((1S,2R,4R)—N-((3-chloro-4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)pyridin-3-yl)acrylate,-   methyl    (E)-3-(5-(N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl)benzamido)pyridin-3-yl)acrylate,-   methyl    (E)-3-(3-(N-((4′-(dimethylamino)-3-fluoro-[1,1′-biphenyl]-4-yl)methyl)benzamido)phenyl)acrylate,-   methyl    (E)-3-(3-(N-((3-chloro-4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl)benzamido)phenyl)acrylate,-   methyl    (E)-3-(3-(N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)benzamido)phenyl)acrylate,-   methyl    (E)-3-(5-(N((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)benzamido)pyridin-3-yl)acrylate,-   methyl    (E)-3-(3-(N-((3-chloro-4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)benzamido)-5-fluorophenyl)acrylate,-   methyl    (E)-3-(3-fluoro-5-(N-((4-(1-methyl-1H-indazol-5-yl)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (E)-3-(3-fluoro-5-(N-(4-(1-methyl-1H-indazol-5-yl)benzyl)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (E)-3-(3-(N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl)benzamido)-5-fluorophenyl)acrylate,-   methyl    (E)-3-(3-(N-(2-chloro-4-(1-methyl-1H-indazol-5-yl)benzyl)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (E)-3-(5-((1S,2R,4R)—N-(2-fluoro-4-(1-methyl-1H-indazol-5-yl)benzyl)bicyclo[2.2.1]heptane-2-carboxamido)pyridin-3-yl)acrylate,-   methyl    (E)-3-(3-(N-((2-fluoro-4-(1-methyl-1H-indazol-5-yl)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (E)-3-(5-(N-((3-chloro-4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)benzamido)pyridin-3-yl)acrylate,-   methyl    (E)-3-(3-(N-(2-chloro-4-(1-methyl-1H-indazol-5-yl)benzyl)cyclohexanecarboxamido)-5-fluorophenyl)acrylate,-   methyl    (E)-3-(5-(N-((2-chloro-4-(1-methyl-1H-indazol-5-yl)phenyl)methyl-d)cyclohexanecarboxamido)pyridin-3-yl)    acrylate,-   methyl    (E)-3-(3-((1S,2R,4R)—N-((2-chloro-4-(1-methyl-1H-indazol-5-yl)phenyl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)-5-fluorophenyl)acrylate,-   methyl    (E)-3-(3-(N-(2-chloro-4-(1-methyl-1H-indazol-5-yl)benzyl)benzamido)-5-fluorophenyl)acrylate,-   methyl    (E)-3-(5-(N-(2-chloro-4-(1-methyl-1H-indazol-5-yl)benzyl)benzamido)pyridin-3-yl)acrylate,-   methyl    (E)-3-(3-fluoro-5-(N-(2-fluoro-4-(1-methyl-1H-indazol-5-yl)benzyl)benzamido)phenyl)acrylate,-   methyl    (E)-3-(3-(N-((2-chloro-4-(1-methyl-1H-indazol-5-yl)phenyl)methyl-d)benzamido)phenyl)acrylate,-   methyl    (E)-3-(5-((1S,2R,4R)—N-((4-(1-methyl-1H-indazol-5-yl)phenyl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)pyridin-3-yl)acrylate,-   methyl    (E)-3-(3-(N-(2-chloro-4-(1-methyl-1H-indazol-5-yl)benzyl)benzamido)phenyl)acrylate,-   methyl    (E)-3-(3-(N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (E)-3-(5-(N-((2-chloro-4-(1-methyl-1H-indazol-5-yl)phenyl)methyl-d)benzamido)pyridin-3-yl)acrylate,-   methyl    (E)-3-(3-(N-((3-chloro-4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (E)-3-(3-(N-((4′-(dimethylamino)-3-fluoro-[1,1′-biphenyl]-4-yl)methyl-d)benzamido)phenyl)acrylate,-   methyl    (E)-3-(5-(N-((4′-(dimethylamino)-3-fluoro-[1,1′-biphenyl]-4-yl)methyl)cyclohexanecarboxamido)pyridin-3-yl)    acrylate,-   methyl    (E)-3-(5-((1S,2R,4R)—N-((3-chloro-4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl)bicyclo[2.2.1]heptane-2-carboxamido)pyridin-3-yl)acrylate,-   methyl    (E)-3-(3-fluoro-5-((1S,2R,4R)—N-((4-(1-methyl-1H-indazol-5-yl)phenyl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)phenyl)acrylate,-   methyl    (E)-3-(3-((1S,2R,4R)—N-((4-(1-methyl-1H-indazol-5-yl)phenyl)methyl-)bicyclo[2.2.1]heptane-2-carboxamido)phenyl)acrylate,-   methyl    (E)-3-(5-(N-((3-chloro-4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl)benzamido)pyridin-3-yl)acrylate,-   methyl    (E)-3-(3-((1S,2R,4R)—N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl)bicyclo[2.2.1]heptane-2-carboxamido)-5-fluorophenyl)acrylate,-   methyl    (E)-3-(3-((1S,2R,4R)—N-((2-fluoro-4-(1-methyl-1H-indazol-5-yl)phenyl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)phenyl)acrylate,-   methyl    (E)-3-(3-(N-((4′-(dimethylamino)-3-fluoro-[1,1′-biphenyl]-4-yl)methyl-d)cyclohexanecarboxamido)-5-fluorophenyl)acrylate,-   methyl    (E)-3-(3-(N-((3-chloro-4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl)benzamido)-5-fluorophenyl)acrylate,-   methyl    (E)-3-(3-((1S,2R,4R)—N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)phenyl)acrylate,-   methyl    (E)-3-(3-((1S,2R,4R)—N-((4′-(dimethylamino)-3-fluoro-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)-5-fluorophenyl)acrylate,-   methyl    (E)-3-(3-(N-((4′-(dimethylamino)-3-fluoro-[1,1′-biphenyl]-4-yl)methyl-d)benzamido)-5-fluorophenyl)acrylate,-   methyl    (E)-3-(3-(N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)benzamido)-5-fluorophenyl)acrylate,-   methyl    (E)-3-(3-((1S,2R,4R)—N-((3-chloro-4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)-5-fluorophenyl)acrylate,-   methyl    (E)-3-(5-(N-(4-(1-methyl-1H-indazol-5-yl)benzyl)benzamido)pyridin-3-yl)acrylate,-   methyl    (E)-3-(5-(N-((4-(1-methyl-1H-indazol-5-yl)phenyl)methyl-d)benzamido)pyridin-3-yl)acrylate,-   methyl    (E)-3-(3-(N-((3-chloro-4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl)cyclohexanecarboxamido)-5-fluorophenyl)acrylate,-   methyl    (E)-3-(5-(N-((4′-(dimethylamino)-3-fluoro-[1,1′-biphenyl]-4-yl)methyl-d)benzamido)pyridin-3-yl)acrylate,-   methyl    (E)-3-(5-((1S,2R,4R)—N-((4′-(dimethylamino)-3-fluoro-[1,1′-biphenyl]-4-yl)methyl)bicyclo[2.2.1]heptane-2-carboxamido)pyridin-3-yl)acrylate,-   methyl    (E)-3-(3-(N-(4-(2-(tert-butoxy)-2-oxoethoxy)benzyl)cyclohexanecarboxamido)phenyl)acrylate,-   tert-butyl    (E)-2-(4-((N-(3-(3-methoxy-3-oxoprop-1-en-1-yl)phenyl)cyclohexanecarboxamido)methyl)phenyl)cyclopropane-1-carboxylate,-   methyl    (E)-3-(3-(N-(4-((2-oxotetrahydro-2H-pyran-3-yl)methyl)benzyl)cyclohexanecarboxamido)phenyl)acrylate,-   cyclobutyl    (E)-3-(4-((N-(3-((E)-3-methoxy-3-oxoprop-1-en-1-yl)phenyl)cyclohexanecarboxamido)methyl)phenyl)acrylate,-   methyl    (E)-3-(3-(N-(4-(6-methoxypyridin-3-yl)benzyl)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (E)-3-(3-(N-((5-(4-(dimethylamino)phenyl)pyridin-2-yl)methyl)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (E)-3-(3-(N-((4-(4-(dimethylamino)phenyl)piperazin-1-yl)methyl)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (E)-3-(3-(N-(4-(3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)benzyl)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (E)-3-(3-(N-(4-(2H-benzo[b][1,4]oxazin-7-yl)benzyl)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (E)-3-(3-(N-(4-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)benzyl)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (E)-3-(3-(N-(4-(quinoxalin-6-yl)benzyl)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (E)-3-(3-(N-(4-(1,2,3,4-tetrahydroquinoxalin-6-yl)benzyl)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (E)-3-(3-(N-(4-(3,4-dihydro-2H-benzo[b][1,4]thiazin-7-yl)benzyl)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (E)-3-(3-(N-(4-(1,1-dioxido-3,4-dihydro-2H-benzo[b][1,4]thiazin-7-yl)benzyl)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (E)-3-(3-(N-(4-(1,1,4,4-tetraoxido-2,3-dihydrobenzo[b][1,4]dithiin-6-yl)benzyl)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    2-(3-(N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl)cyclohexanecarboxamido)phenyl)cyclopropane-1-carboxylate,-   methyl    2-(3-(N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl)cyclohexanecarboxamido)phenyl)cyclobutane-1-carboxylate,-   methyl    2-(3-(N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl)cyclohexanecarboxamido)phenyl)cyclopentane-1-carboxylate,-   methyl    2-(3-(N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl)cyclohexanecarboxamido)phenyl)cyclohexane-1-carboxylate,-   methyl    (E)-3-(3-(N-(4-(benzo[d]oxazol-6-yl)benzyl)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (E)-3-(3-(N-(4-(1H-benzo[d]imidazol-6-yl)benzyl)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (E)-3-(3-(N-(4-(benzo[d]thiazol-6-yl)benzyl)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (E)-3-(3-(N-(4-(2-methylbenzo[d]thiazol-6-yl)benzyl)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (E)-3-(3-(N-(4-(2-methylbenzo[d]oxazol-6-yl)benzyl)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (E)-3-(3-(N-(4-(2-methyl-1H-benzo[d]imidazol-6-yl)benzyl)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (E)-3-(3-(N-(4-(1,2-dimethyl-1H-benzo[d]imidazol-6-yl)benzyl)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (E)-3-(3-(N-(4-(1-methyl-1H-benzo[d]imidazol-6-yl)benzyl)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (E)-3-(3-(N-(4-(benzo[d]isoxazol-5-yl)benzyl)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (E)-3-(3-(N-(4-(benzo[d]isothiazol-5-yl)benzyl)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    3-(3-(N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl)bicyclo[2.2.1]heptane-2-carboxamido)phenyl)propanoate,-   methyl    2-(3-(N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl)bicyclo[2.2.1]heptane-2-carboxamido)phenoxy)acetate,-   3-(N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl)bicyclo[2.2.1]heptane-2-carboxamido)benzyl    acetate,-   N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl)-N-(3-((2-oxotetrahydro-2H-pyran-3-yl)methyl)phenyl)cyclohexanecarboxamide,-   N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl)-N-(3-((2-oxotetrahydro-2H-pyran-4-yl)methyl)phenyl)cyclohexanecarboxamide,-   methyl    (E)-3-(3-(N-((4-(2-(tert-butoxy)-2-oxoethoxy)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate,-   tert-butyl (E)-2-(4-((N-(3-(3 -methoxy-3    -oxoprop-1-en-1-yl)phenyl)cyclohexanecarboxamido)methyl-d)phenyl)cyclopropane-1-carboxylate,-   methyl    (E)-3-(3-(N-((4-((2-oxotetrahydro-2H-pyran-3-yl)methyl)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate,-   cyclobutyl    (E)-3-(4-((N-(3-((E)-3-methoxy-3-oxoprop-1-en-1-yl)phenyl)cyclohexanecarboxamido)methyl-d)phenyl)acrylate,-   methyl    (E)-3-(3-(N-((4-(6-methoxypyridin-3-yl)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (E)-3-(3-(N-((5-(4-(dimethylamino)phenyl)pyridin-2-yl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (E)-3-(3-(N-((4-(4-(dimethylamino)phenyl)piperazin-1-yl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (E)-3-(3-(N-((4-(3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (E)-3-(3-(N4(4-(2H-benzo[b][1,4]oxazin-7-yl)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (E)-3-(3-(N-((4-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (E)-3-(3-(N-((4-(quinoxalin-6-yl)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (E)-3-(3-(N-((4-(1,2,3,4-tetrahydroquinoxalin-6-yl)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (E)-3-(3-(N-((4-(3,4-dihydro-2H-benzo[b][1,4]thiazin-7-yl)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (E)-3-(3-(N-((4-(1,1-dioxido-3,4-dihydro-2H-benzo[b][1,4]thiazin-7-yl)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (E)-3-(3-(N-((4-(1,1,4,4-tetraoxido-2,3-dihydrobenzo[b][1,4]dithiin-6-yl)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    2-(3-(N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)cyclohexanecarboxamido)phenyl)cyclopropane-1-carboxylate,-   methyl    2-(3-(N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)cyclohexanecarboxamido)phenyl)cyclobutane-1-carboxylate,-   methyl    2-(3-(N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)cyclohexanecarboxamido)phenyl)cyclopentane-1-carboxylate,-   methyl    2-(3-(N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)cyclohexanecarboxamido)phenyl)cyclohexane-1-carboxylate,-   methyl    (E)-3-(3-(N-((4-(benzo[d]oxazol-6-yl)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (E)-3-(3-(N-((4-(1H-benzo[d]imidazol-6-yl)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (E)-3-(3-(N-((4-(benzo[d]thiazol-6-yl)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (E)-3-(3-(N-((4-(2-methylbenzo[d]thiazol-6-yl)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (E)-3-(3-(N-((4-(2-methylbenzo[d]oxazol-6-yl)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (E)-3-(3-(N-((4-(2-methyl-1H-benzo[d]imidazol-6-yl)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (E)-3-(3-(N-((4-(1,2-dimethyl-1H-benzo[d]imidazol-6-yl)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (E)-3-(3-(N-((4-(1-methyl-1H-benzo[d]imidazol-6-yl)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (E)-3-(3-(N-((4-(benzo[d]isoxazol-5-yl)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (E)-3-(3-(N-((4-(benzo[d]isothiazol-5-yl)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    3-(3-(N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)phenyl)propanoate,-   methyl    2-(3-(N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)phenoxy)acetate,-   3-(N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)benzyl    acetate,-   N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)-N-(3-((2-oxotetrahydro-2H-pyran-3-yl)methyl)phenyl)cyclohexanecarboxamide,-   N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)-N-(3-((2-oxotetrahydro-2H-pyran-4-yl)methyl)phenyl)cyclohexanecarboxamide,-   methyl    (E)-3-(3-((1S,2R,4R)—N—((S)-(4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)-5-fluorophenyl)acrylate,-   methyl    (E)-3-(3-((1S,2R,4R)—N—((R)-(4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)-5-fluorophenyl)acrylate,-   methyl    (E)-3-(3-((1S,2S,4R)—N—((S)-(4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)-5-fluorophenyl)acrylate,-   methyl    (E)-3-(3-((1S,2S,4R)—N—((R)-(4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)-5-fluorophenyl)acrylate,-   methyl    (E)-3-(3-((1R,2S,4S)—N—((S)-(4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)-5-fluorophenyl)acrylate,-   methyl    (E)-3-(3-((1R,2S,4S)—N—((R)-(4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)-5-fluorophenyl)acrylate,-   methyl    (E)-3-(3-((1R,2R,4S)—N-((5)-(4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)-5-fluorophenyl)acrylate,-   methyl    (E)-3-(3-((1R,2R,4S)—N—((R)-(4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)-5-fluorophenyl)acrylate,-   methyl    (E)-3-(3-((1S,2R,4R)—N-((5)-(4′-(dimethylamino)-3-fluoro-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)-5-fluorophenyl)acrylate,-   methyl    (E)-3-(3-((1S,2R,4R)—N—((R)-(4′-(dimethylamino)-3-fluoro-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)-5-fluorophenyl)acrylate,-   methyl    (E)-3-(3-((1S,2S,4R)—N—((S)-(4′-(dimethylamino)-3-fluoro-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)-5-fluorophenyl)acrylate,-   methyl    (E)-3-(3-((1S,2S,4R)—N—((R)-(4′-(dimethylamino)-3-fluoro-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)-5-fluorophenyl)acrylate,-   methyl    (E)-3-(3-((1R,2S,4S)—N—((S)-(4′-(dimethylamino)-3-fluoro-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)-5-fluorophenyl)acrylate,-   methyl    (E)-3-(3-((1R,2S,4S)—N—((R)-(4′-(dimethylamino)-3-fluoro-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)-5-fluorophenyl)acrylate,-   methyl    (E)-3-(3-((1R,2R,4S)—N—((S)-(4′-(dimethylamino)-3-fluoro-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)-5-fluorophenyl)acrylate,-   methyl    (E)-3-(3-((1R,2R,4S)—N—((R)-(4′-(dimethylamino)-3-fluoro-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)-5-fluorophenyl)acrylate,-   methyl    (R,E)-3-(3-(N-((4′-(dimethylamino)-3-fluoro-[1,1′-biphenyl]-4-yl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (S,E)-3-(3-(N-((4′-(dimethylamino)-3-fluoro-[1,1′-biphenyl]-4-yl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (R,E)-3-(3-(N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (S,E)-3-(3-(N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (R,E)-3-(3-fluoro-5-(N-((4-(1-methyl-1H-indazol-5-yl)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (S,E)-3-(3-fluoro-5-(N-((4-(1-methyl-1H-indazol-5-yl)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (R,E)-3-(3-(N-((2-fluoro-4-(1-methyl-1H-indazol-5-yl)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate,    or-   methyl    (S,E)-3-(3-(N-((2-fluoro-4-(1-methyl-1H-indazol-5-yl)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate.

In particular working embodiments, the compound is selected from

-   methyl    (E)-3-(3-fluoro-5-(N-((4-(1-methyl-1H-indazol-5-yl)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate

-   methyl    (E)-3-(3-(N-((2-fluoro-4-(1-methyl-1H-indazol-5-yl)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate

-   methyl    (E)-3-(3-((1R,4S)—N-((4′-(dimethylamino)-3-fluoro-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)-5-fluorophenyl)acrylate

-   methyl    (E)-3-(3-((1R,4S)—N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)-5-fluorophenyl)acrylate,    and

-   methyl    (E)-3-(3-(N-((4′-(dimethylamino)-3-fluoro-[1,1′-biphenyl]-4-yl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate

In other particular embodiments, the compound is

-   methyl    (E)-3-(3-((1S,2R,4R)—N—((S)-(4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)-5-fluorophenyl)acrylate,-   methyl    (E)-3-(3-((1S,2R,4R)—N—((R)-(4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)-5-fluorophenyl)acrylate,-   methyl    (E)-3-(3-((1S,2S,4R)—N—((S)-(4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)-5-fluorophenyl)acrylate,-   methyl    (E)-3-(3-((1S,2S,4R)—N—((R)-(4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)-5-fluorophenyl)acrylate,-   methyl    (E)-3-(3-((1R,2S,4S)—N—((S)-(4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)-5-fluorophenyl)acrylate,-   methyl    (E)-3-(3-((1R,2S,4S)—N—((R)-(4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)-5-fluorophenyl)acrylate,-   methyl    (E)-3-(3-((1R,2R,4S)—N—((S)-(4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)-5-fluorophenyl)acrylate,-   methyl    (E)-3-(3-((1R,2R,4S)—N—((R)-(4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)-5-fluorophenyl)acrylate,-   methyl    (E)-3-(3-((1S,2R,4R)—N—((S)-(4′-(dimethylamino)-3-fluoro-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)-5-fluorophenyl)acrylate,-   methyl    (E)-3-(3-((1S,2R,4R)—N—((R)-(4′-(dimethylamino)-3-fluoro-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)-5-fluorophenyl)acrylate,-   methyl    (E)-3-(3-((1S,2S,4R)—N—((S)-(4′-(dimethylamino)-3-fluoro-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)-5-fluorophenyl)acrylate,-   methyl    (E)-3-(3-((1S,2S,4R)—N—((R)-(4′-(dimethylamino)-3-fluoro-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)-5-fluorophenyl)acrylate,-   methyl    (E)-3-(3-((1R,2S,4S)—N—((S)-(4′-(dimethylamino)-3-fluoro-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)-5-fluorophenyl)acrylate,-   methyl    (E)-3-(3-((1R,2S,4S)—N—((R)-(4′-(dimethylamino)-3-fluoro-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)-5-fluorophenyl)acrylate,-   methyl    (E)-3-(3-((1R,2R,4S)—N—((S)-(4′-(dimethylamino)-3-fluoro-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)-5-fluorophenyl)acrylate,-   methyl    (E)-3-(3-((1R,2R,4S)—N—((R)-(4′-(dimethylamino)-3-fluoro-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)-5-fluorophenyl)acrylate,-   methyl    (R,E)-3-(3-(N-((4′-(dimethylamino)-3-fluoro-[1,1′-biphenyl]-4-yl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (S,E)-3-(3-(N-((4′-(dimethylamino)-3-fluoro-[1,1′-biphenyl]-4-yl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (R,E)-3-(3-(N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (S,E)-3-(3-(N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (R,E)-3-(3-fluoro-5-(N-((4-(1-methyl-1H-indazol-5-yl)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (S,E)-3-(3-fluoro-5-(N-((4-(1-methyl-1H-indazol-5-yl)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate,-   methyl    (R,E)-3-(3-(N-((2-fluoro-4-(1-methyl-1H-indazol-5-yl)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate,    or-   methyl    (S,E)-3-(3-(N-((2-fluoro-4-(1-methyl-1H-indazol-5-yl)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate.

Also provided herein are kits that include any FXR agonist (orcomposition containing such an agonist) described herein and a devicefor localized delivery within a region of the intestines, such as theileum or colon. In certain embodiments, the device is a syringe, bag, ora pressurized container.

IV. COMPOSITIONS

Also disclosed herein are pharmaceutical compositions comprising atleast one compound having formulas 1-18. Remington's PharmaceuticalSciences, by E. W. Martin, Mack Publishing Co., Easton, Pa., 15thEdition, 1975, incorporated herein by reference, describes exemplaryformulations (and components thereof) suitable for pharmaceuticaldelivery of the disclosed compounds. Pharmaceutical compositionscomprising at least one of the disclosed compounds can be formulated foruse in human or veterinary medicine. Particular formulations of adisclosed pharmaceutical composition may depend, for example, on themode of administration (e.g., oral). In some embodiments, disclosedpharmaceutical compositions include a pharmaceutically acceptablecarrier in addition to at least one or two or more active ingredients,such as a compound or compounds disclosed herein. In other embodiments,other medicinal or pharmaceutical agents, for example, with similar,related or complementary effects on the affliction being treated (suchas obesity, dyslipidemia, or diabetes), can also be included as activeingredients in a pharmaceutical composition. For example, one or more ofthe disclosed compounds can be formulated with one or more of (such as1, 2, 3, 4, or 5 of) an antibiotic (e.g., metronidazole, vancomycin,and/or fidaxomicin), statin, alpha-glucosidase inhibitor, amylinagonist, dipeptidyl-peptidase 4 (DPP-4) inhibitor (such as sitagliptin,vildagliptin, saxagliptin, linagliptin, anaglptin, teneligliptin,alogliptin, gemiglptin, or dutoglpitin), meglitinide, sulfonylurea,peroxisome proliferator-activated receptor (PPAR)-gamma agonist (e.g., athiazolidinedione (TZD) [such as ioglitazone, rosiglitazone,rivoglitazone, or troglitazone], aleglitazar, farglitazar, muraglitazar,or tesaglitazar), anti-inflammatory agent (e.g., oral corticosteroid),chemotherapeutic, biologic, radiotherapeutic, nicotinamideribonucleoside, analogs of nicotinamide ribonucleoside that promotesNAD+ production of which is a substrate for many enzymatic reactionssuch as p450s which are a target of FXR (e.g., see Yang et al., J. Med.Chem. 50:6458-61, 2007), and the like.

Pharmaceutically acceptable carriers useful for the disclosed method andcomposition will depend on the particular mode of administration beingemployed. For example, for solid compositions (e.g., powder, pill,tablet, or capsule forms), conventional non-toxic solid carriers caninclude, without limitation, pharmaceutical grades of sugars, such asmannitol or lactose, polysaccharides, such as starch, or salts oforganic acids, such as magnesium stearate. In addition to biologicallyneutral carriers, pharmaceutical compositions can optionally containamounts of auxiliary substances (e.g., excipients), such as wetting oremulsifying agents, preservatives, and pH buffering agents and the like;for example, sodium acetate or sorbitan monolaurate. Other non-limitingexcipients include nonionic solubilizers, such as cremophor, orproteins, such as human serum albumin or plasma preparations. In someembodiments, the pharmaceutical composition comprises a sufficientamount of a disclosed compound to have a desired therapeutic effect.Typically, the disclosed compound constitutes greater than 0% to lessthan 100% of the pharmaceutical composition, such as 10% or less, 20% orless, 30% or less, 40% or less, 50% or less, 60% or less, 70% or less,80% or less, 90% or less, or 90% to less than 100% of the pharmaceuticalcomposition.

The disclosed pharmaceutical compositions may be formulated as apharmaceutically acceptable salt, solvate, hydrate, N-oxide orcombination thereof, of a disclosed compound. Additionally, thepharmaceutical composition may comprise one or more polymorph of thedisclosed compound. Pharmaceutically acceptable salts are salts of afree base form of a compound that possesses the desired pharmacologicalactivity of the free base. These salts may be derived from inorganic ororganic acids. Non-limiting examples of suitable inorganic acids includehydrochloric acid, nitric acid, hydrobromic acid, sulfuric acid,hydriodic acid, and phosphoric acid. Non-limiting examples of suitableorganic acids include acetic acid, propionic acid, glycolic acid, lacticacid, pyruvic acid, malonic acid, succinic acid, malic acid, maleicacid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamicacid, mandelic acid, methanesulfonic acid, ethanesulfonic acid,p-toluenesulfonic acid, methyl sulfonic acid, salicylic acid, formicacid, trichloroacetic acid, trifluoroacetic acid, gluconic acid,asparagic acid, aspartic acid, benzenesulfonic acid, p-toluenesulfonicacid, naphthalenesulfonic acid, and the like. Examples of other suitablepharmaceutically acceptable salts are found in Remington'sPharmaceutical Sciences, 17th Edition, Mack Publishing Company, Easton,Pa., 1985.

In some embodiments, the compounds disclosed herein may be formulated tohave a suitable particle size. A suitable particle size may be one whichreduces or substantially precludes separation of the components of thecomposition, e.g., no separation between the drug and any othercomponents of the composition, such as a second drug, a pharmaceuticallyacceptable excipient, a corticosteroid, an antibiotic or any combinationthereof. Additionally, the particle size may be selected to ensure thecomposition is suitable for delivery, such as oral delivery.

In certain embodiments, the composition further includes an entericcoating. Typically, an enteric coating is a polymer barrier applied toan oral medication to help protect the drug from the acidity and/orenzymes of the stomach, esophagus and/or mouth. In some embodiments,this coating can reduce or substantially prevent systemic delivery ofthe disclosed compound, thereby allowing substantially selectivedelivery to the intestines. In some embodiments, the enteric coatingwill not dissolve in the acid environment of the stomach, which has anacidic, pH of about 3, but will dissolve in the alkaline environments ofthe small intestine, with, for example, a pH of about 7 to 9. Materialsused for enteric coating include, but are not limited to, fatty acids,waxes, shellac, plastics and plant fibers. In some embodiments, thecoating may comprise methyl acrylate-methacrylic acid copolymers,cellulose acetate succinate, hydroxy propyl methyl cellulose phthalate,hydroxy propyl methyl cellulose acetate succinate (hypromellose acetatesuccinate), polyvinyl acetate phthalate (PVAP), methylmethacrylate-methacrylic acid copolymers, shellac, cellulose acetatetrimellitate, sodium alginate, or any combination thereof.

V. METHODS OF MAKING THE COMPOUNDS

Embodiments of a method of making compounds that have formulas 1-18 arealso disclosed herein. A general method of making the compoundscomprises reacting an aldehyde with a first amine to form an imine,reacting the imine with a reducing agent to form a second amine, andreacting the second amine with an activated carboxylic acid derivativeor a carboxylic acid to form an amide.

Other embodiments further comprise contacting the aldehyde with aboronic acid, contacting the amide with a vinyl ester, contacting thefirst amine with a vinyl ester, contacting the amide with a boronicacid, or any combination thereof. In certain embodiments the reducingagent is a deuterated reducing agent, and the compound comprisesdeuterium.

One exemplary embodiment of the general method is shown in Scheme 1.

A. Vinyl group coupling

With reference to scheme 1, a protected aromatic amine 2 was coupled toa vinyl ester 4 by a suitable coupling technique to form compound 6. Theamine of the aromatic amine 2 was protected by a suitable protectinggroup, as will be understood by a person of ordinary skill in the art.Additional information concerning protecting groups is provided byGreene and Wuts, Protective Groups in Organic Synthesis; 3rd Ed.; JohnWiley & Sons, New York, 1999, which is incorporated herein by reference.Exemplary amine protecting groups include, but are not limited to,tert-butyloxycarbonyl (Boc), benzyl, benzoyl, or benzoyloxycarbonyl(Cbz). In some embodiments the technique is a Stille coupling. Incertain working embodiments, coupling comprised treating the protectedaromatic amine with a vinyl group in the presence of a suitablecatalyst, such as a palladium catalyst, and optionally, a suitablephosphine compound. Suitable palladium catalysts include, but are notlimited to, Bis(dibenzylideneacetone)palladium (Pd2(dba)₃) or palladiumacetate (Pd(OAc)₃). In certain working examples Pd₂(dba)₃ was used as acatalyst with tri(o-tolyl)phosphine (P(o-tol)₃) as the phosphine. Thecoupling reaction is conducted in any suitable solvent, such asdimethylformamide, at a temperature effective to facilitate a reaction.In some embodiments the effective temperature is from greater than 0° C.to at least 130° C., such as from about 20° C. to about 110° C., fromabout 80° C. to about 100° C. In certain working embodiments thetemperature was about 95° C.

B. Reductive Amination

The amine protecting group of compound 6 was removed by treatment with asuitable reagent. Suitable de-protection reagents and conditions for aspecific protecting group can be selected by a person of ordinary skillin the art, and is further disclosed by consulting Greene and Wuts. Incertain working embodiments, trifluoroacetic acid (TFA) was used toremove a Boc protecting group. In certain disclosed embodiments, thede-protected amine (not shown) was then treated with an aldehyde, suchas aldehyde 8, in the presence of a reducing agent. In otherembodiments, the amine was treated with an aldehyde, and subsequentlytreated by a reducing agent. The reducing agent is selected to place adesired R⁴ group into the molecule. In some embodiments R⁴ is hydrogen;in others it is deuterium. Suitable reducing agents include, but are notlimited to, sodium triacetoxyborohydride, sodiumtriacetoxyborodeuteride, sodium cyanoborohydride, sodiumcyanoborodeuteride, sodium borohydride, lithium borohydride, sodiumborodeuteride or lithium borodeuteride. Suitable solvents for thereduction include, but are not limited to, toluene, halogenatedsolvents, THF, hexanes, cyclohexane, acetic acid, deuterated aceticacid, alcohols such as methanol, ethanol propanol, isopropanol, ordeuterated alcohols such as methanol-d₄. Typically, the reducing agentwas NaBH(OAc)₃, NaBD(OAc)₃, NaBD₃CN, NaBH₄ or NaBD₄ and the solvent wasTHF, CD₃OD, acetic acid or deuterated acetic acid.

C. Acylation

Subsequent to the reductive amination, compound 10 was acylated withacylating agent 12 under suitable conditions, such as by treatment witha carboxylic acid or an activated carboxylic acid derivative, such as anacid chloride, an acid bromide, or an anhydride. A person of ordinaryskill in the art will understand which activated carboxylic acidderivatives are suitable for a particular carboxylic acid.Alternatively, a carboxylic acid may be coupled to the amine using asuitable coupling reagent known to a person of ordinary skill in theart. Exemplary coupling reagents include, but are not limited to, HATU,dicyclohexylcarbodiimide (DCCI, DCC) or1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC, EDCI, EDAC). Inworking embodiments the carboxylic acid was activated by forming an acidchloride. The acylation reactions proceed in a suitable solvent,typically an aprotic solvent, such as pyridine, dichloromethane,chloroform, dioxane, toluene, DMF, THF or acetonitrile. Optionally, thereaction with a carboxylic acid or a carboxylic acid derivative mayproceed in the presence of one or more additional compounds, such aspotassium carbonate, triethylamine, diisopropylethylamine, sodiumcarbonate, 4-(dimethylamino)pyridine (DMAP) or pyridine. The reactionsare performed at a temperature effective to facilitate the reaction,such as from greater than about −10° C. to greater than about 120° C.,typically from about 5° C. to about 90° C., more typically from about25° C. to about 65° C.

D. Boronic Acid Coupling

After the acylation reaction, compound 14 was treated with a boronicacid 16, in a Suzuki-type coupling. In some embodiments, the couplingwas performed in the presence of a catalyst effective to facilitate thecoupling reaction, and optionally in the presence of one or moreadditional compounds. Typical catalysts for a Suzuki coupling arepalladium or nickel catalysts, including but not limited to,NiCl₂(dppf), NiCl₂(dppp), Pd(PPh₃)₄, Pd(OAC)₂ or PdCl₂(PPh₃)₄. Inworking embodiments the catalyst was Pd(PPh₃)₄. Typical additionalcompounds include, but are not limited to, triphenylphosphine (PPh₃),and/or bases such as potassium carbonate, sodium carbonate, cesiumcarbonate, sodium hydroxide, potassium hydroxide, triethylamine, sodiumethoxide, sodium methoxide, tripotassium phosphate or any combinationthereof. In certain working embodiments, the additional compound wassodium carbonate. The coupling reaction is performed in any suitablesolvent, such as DMF, ethanol, methanol, isopropanol, propanol, benzene,toluene, THF, dioxane, water or any combination thereof. In certainworking embodiments, DMF-ethanol-water was used as the solvent.

A person of ordinary skill in the art will recognize that the varioussteps described above with reference to Scheme 1 do not necessarily haveto be performed in the particular order depicted. The reactions can beperformed in any order suitable to result in making the desired compound18. For example, in certain embodiments, the sequence of reactionsfollowed the order described in Scheme 2.

With reference to Scheme 2, boronic acid 16 was first coupled toaldehyde 8. The resulting product 20 was then treated with an aminecompound 22 in a reductive amination step to form compound 24. Compound24 was then acylated using acylating reagent 12 to form compound 26,which was then coupled to vinyl ester 4 to form compound 18.

Another variation of Scheme 1 is shown in Scheme 3. In this scheme, thecompounds are made using a solid-phase synthetic method as used for thesynthesis of fexaramine in U.S. Pat. No. 7,647,217, which isincorporated herein by reference. Thus, protected amine 6, where R ishydrogen, is immobilized onto a solid support 34, such as a bead orresin, typically Merrifield resin, through the action of a suitablebase, for example, cesium carbonate, sodium carbonate or potassiumcarbonate, to make conjugate 36. The reductive amination, acylation andboronic acid coupling steps then proceed on the immobilized compound asdescribed for Scheme 1, making conjugates 38, 40 and 42 respectively.Conjugate 42 is then treated with an alkoxide salt, such as sodiummethoxide, to release the desired compound 18 from the solid support.

In certain embodiments, an alternative reaction pathway was followed.Scheme 4 outlines an exemplary alternative route. First, a halogenatednitrobenzene 28 was coupled to vinyl ester 4 in the presence of asuitable catalyst and an additional compound (not shown) to formcompound 30. In working embodiments the catalyst was Pd(OAc)₂, and theadditional compound was sodium acetate. The nitro group of compound 30was then reduced to an amine by a suitable reagent to form amine 32.Suitable reagents include, but are not limited to, tin chloride, ironpowder in an acid medium, zinc powder or catalytic hydrogenation using atransition metal catalyst comprising palladium, platinium, nickel,rhodium or ruthenium. In working embodiments tin chloride (SnCl₂) wasused as the reducing agent.

Amine 32 was then treated with an aldehyde 8 in a de-hydration reaction.Suitable dehydrating agents include, but are not limited to, an acidcatalyst such as para-toluene sulfonic acid, a base such astriethylamine, malononitrile, molecular sieve, magnesium sulfate, sodiumsulfate, or any combination thereof. Suitable solvents for thede-hydration reaction include toluene, xylenes, DMSO, DMF, THF, alcoholssuch as methanol or any combination thereof. Resulting imine compound 34was then treated with a suitable reducing agent to form amine 10. Inworking embodiments, a deuterated reducing agent was used. In someembodiments sodium cyanodeuteroborohydride was used, and in otherssodium deuteroborohydride was used. Any suitable, non-protonated solventcan be used, and in some working embodiments the solvent was methanol-d₄and in others it was THF.

Amine 10 was then treated with acylating reagent 12, as described abovewith reference to Scheme 1, to form compound 14. In working embodimentsthe amine was treated with carboxylic acids in the presence of HATU anddiisopropylethylamine in DMF. In other working embodiments the amine wastreated with carboxylic acid chlorides in dichloromethane in thepresence of triethylamine.

Compound 14 was then treated with boronic acid 16 as described abovewith reference to Scheme 1. In certain working embodiments, compound 14was treated with boronic acid 16 in a DME-ethanol-water solvent system,in the presence of Pd(PPh₃)₄ and sodium carbonate. In other workingembodiments dioxane-water was used as the solvent, Pd(dppf)Cl₂ was thecatalyst and potassium carbonate was used as a base.

One exemplary method of making compounds having formula 13 is shown inScheme 5. This method is a modification of the method of Lee andHartwig, J. Org. Chem. 2001, 66, 3402-3415.

With reference to Scheme 5, compound 20 is reacted with a catalyst 22and a base 24 in a suitable solvent, to form compound 26. Leaving groupLG on compound 20 is any suitable leaving group, such as a halide,mesylate, tosylate or trifluoromethylsulfonate. Catalyst 22 is anycatalyst that facilitates the formation of compound 26. Suitablecatalysts include, but are not limited to, palladium catalysts such asPd(OAc)₂, and may also comprise one or more ligands, such as PCy₃, orsterically hindered N-heterocyclic carbine ligands. The amount of thecatalyst used is any suitable amount to catalyze the reaction at asuitable rate, such as from about 1 mol % to greater than about 20 mol%, preferably from about 5 mol % to about 10 mol %. Base 24 is anysuitable base that facilitates the reaction. In some embodiments anexcess of the base is used, such as from greater than 1 equivalent togreater than about 5 equivalents, preferably from about 1.1 equivalentsto about 2 equivalents. Suitable bases include, but are not limited to,tert-butoxide salts, such as sodium, lithium or potassium tert-butoxide.The solvent can be any solvent suitable to facilitate a reaction. Insome embodiments the solvent is 1,4-dioxane.

Embodiments of a method of making prodrugs of compounds having formulas1-13 are also disclosed herein. One general method of making prodrugs isdisclosed by Poon, et al. Bioorg. med. Chem. Lett. 2005, 15: 2259-2263,and is shown in Scheme 6. Briefly, the method comprises making thethioester of the compound, and forming the ortho ester or imidate.

With reference to Scheme 6, ester compound 30 is reacted with reagentsuitable to form thioester compound 32. Suitable reagents include, butare not limited to, Lawesson's reagent or P₂S₅. The reaction isperformed in a suitable solvent, usually an aprotic solvent such astoluene, acetonitrile, cyclohexane, dichloromethane, or chloroform. Thereaction may also be heated, such as to reflux.

The thioester compound 32 is then reacted with reagents suitable to formthe desired prodrug, in the presence of a metal salt and a base. Themetal salt is any metal salt suitable to mediate thedesulfurization-condensation reaction between the thioester compound 32and the alcohol or amine Suitable metal salts include, but are notlimited to, silver salts such as AgoTf. Suitable bases include, but arenot limited to organic bases such as triethylamine ordiisopropylethylamine. The reactions are performed in solvent suitableto facilitate the reaction, such as an aprotic solvent. Suitablesolvents include, but are not limited to, acetonitrile, DMF,dimethylacetyl, N-methyl-2-pyrrolidone.

To form compound 34, thioester 32 is reacted with dibenzylascorbate,AgOTf and triethylamine in acetonitrile. An intermediate compound isformed initially (not shown) which is then reacted with hydrogen in thepresence of a palladium catalyst in alcohol to form compound 34.Compound 36 is formed by reacting compound 32 with hydroxylamine, in thepresence of AgOTf and triethylamine in acetonitrile. The intermediatecompound (not shown) is then reacted with 2-bromoacetic acid and sodiumhydroxide, to form compound 36. Compound 38 is made by reacting compound32 with serine-OMe in the presence of AgOTf and triethylamine, inacetonitrile. The intermediate compound formed (not shown) is thenreacted with Sodium trimethylsilanolate (NaOTMS) in THF to form compound38.

A method of making compounds having formula 17 and 18 is shown in Scheme7, and is a modification of a method disclosed by Ates and Curran, J.Am. Chem. Soc. 2001, 123: 5130-5131.

With reference to Scheme 7, compound 40 is reacted with a methylatingagent, such as methyl trifluoromethanesulfonate, in a suitable solventto make compound 42. Suitable solvents include, but are not limited to,halogenated solvents such as dichloromethane and chloroform. Compound 42is reacted with a metal alkoxide solution, such as sodium methoxide inmethanol, to form compound 44, an exemplary compound satisfying formula17. Compound 44 is further reacted with dimethyl tartrate in a vacuum toform compound 46, an exemplary compound satisfying formula 18.

VI. METHODS OF USING THE COMPOUNDS/COMPOSITIONS

Orally delivered fexaramine (Fex) (Downes et al., Mol Cell 11:1079-1092,2003) is poorly absorbed, resulting in intestinally-restricted FXRactivation. It is shown herein that despite this restricted activation,Fex treatment of diet-induced obesity (DIO) mice produces a novelmetabolic profile that includes reduced weight gain, decreasedinflammation, browning of white adipose tissue and increased insulinsensitization. The beneficial systemic efficacy achieved with Fexsuggests intestinal FXR therapy as a potentially safer approach in thetreatment of insulin resistance and metabolic syndrome.

It is shown herein that the gut-biased FXR agonist fexaramine hasprofound metabolic benefits in a mouse model of obesity. Fex protectsagainst diet-induced weight gain by promoting the expression of genesinvolved in thermogenesis, mitochondrial biogenesis, and fatty acidoxidation. Linked to the unexpected browning of white adipose, Fexlowers inflammatory cytokine levels while up-regulating β-adrenergicsignaling. These changes appear to be mediated in part by a change inbile acid levels and composition. In addition, intestinal-specific FXRactivation corrected numerous obesity-related defects, enhanced glucosetolerance, and lowered hepatic glucose production. Notably, thesephysiologic changes are dependent on FXR expression and result inhepatic insulin sensitization and BAT activation, properties notformerly associated with this class of drug.

The initial event triggering systemic metabolic activation is likelycoordinated by FGF15, a key regulator of energy expenditure reported toincrease metabolic rate, and improve glucose and lipid homeostasiswithout significant changes in food intake (Fu et al., Endocrinology145:2594-2603, 2004; Bhatnagar et al., J Biol Chem 284:10023-10033,2009). The absence of a change in food intake is significant as failureof appetite control is a major reason for weight gain (Foster-Schubert &Cummings, Endocr Rev 27:779-793, 2006). Thus, systemic increases inenergy expenditure, as seen in Fex-treated mice, may offer a viablealternative for obesity treatments. However, this explanation alone isnot sufficient as systemic FXR agonists, while robustly inducing FGF15,do not display many of the benefits of gut-biased FXR activation.

One major difference between gut-biased and systemic FXR activation isthe impact on serum bile acids, which for Fex includes a marked changein the relative composition of circulating BAs. A reduction in hepaticCYP7A1 accompanied by an increase in CYP7B1 expression shifts BAsynthesis away from cholic acid towards chenodeoxycholic acidderivatives, most notably lithocholic acid. While the absolute amount oflithocholic acid did not change following Fex the relative amountincreased dramatically. Lithocholic acid is a hydrophobic secondary bileacid and the most potent endogenous ligand for the G protein-coupledbile acid receptor TGR5 (Ullmer et al., Br. J. Pharmacol. 169:671-684,2013). Interestingly, Fex treatment induces metabolic changes similar tothose observed with systemic administration of a synthetic TGR5 agonist(Ullmer et al., Br. J. Pharmacol. 169:671-684, 2013). Also, induction ofDIO2, a downstream target of TGR5 (Watanabe et al., Nature 439:484-489,2006), in BAT with oral Fex implicates this pathway in the observedincreased energy expenditure. Indeed, the metabolic improvementsattributed to Fex treatment were tempered in TGR^(−/−) mice, indicatingthat TGR5 activation is important in meditating some of the actions ofFex. Furthermore, the coordinate “browning” of the WAT depot provides anindependent yet complementary contribution to increased thermogeniccapacity.

These results uncover a new therapeutic avenue to manipulate energyexpenditure without appetite changes through intestinally-biasedactivation of the nuclear receptor FXR. While contrary indications havebeen recently reported, the integral role of FXR in gut homeostasisconfounds these studies (Kim et al., J Lipid Res 48:2664-2672, 2007; Li,et al., Nat Commun 4:2384, 2013). Gut-restricted drugs such as Fexinherently offer improved safety profiles, achieving systemic efficacywhile avoiding systemic toxicity. In support of the remarkable metabolicimprovements achieved via oral Fex treatment, intestinal FXR has beenrecently identified as a molecular target of vertical sleeve gastrectomy(Ryan et al., Nature 509:183-188, 2014), indicating that Fex may offer anon-surgical alternative for the control of metabolic disease.

A. Treatment or Prevention of Metabolic Disorders

Treatment of subjects, including diet-induced obesity (DIO) subjects,with one or more of the disclosed FXR agonists (such as two or more,three or more, four or more, or five or more of the disclosed FXRagonists, such as 2, 3, 4, or 5 of the disclosed FXR agonists) producesbeneficial body-wide metabolic effects such as reduced weight gain,decreased inflammation, browning of white adipose tissue, activation ofBAT, improved insulin sensitization, or combinations thereof. Thus,intestinally-restricted FXR administration is superior to systemic FXRtherapy for body-wide metabolic disorders including obesity andmetabolic syndrome. One or more of the FXR agonists disclosed herein canbe administered to a gastrointestinal (GI) tract of the subject toactivate FXR receptors in the intestines, and thereby treat or prevent ametabolic disorder in the subject. Thus, the FXR agonist(s) can beadministered to, without limitation, the mouth (such as by injection orby ingestion by the subject), the esophagus, the stomach or theintestines themselves.

Orally delivered, these agonists can in some examples be ineffectivelyabsorbed, resulting in intestinally-restricted FXR activation. In someembodiments, FXR activation is completely limited to the intestine. Insome embodiments, administration of one or more of the disclosedagonists does not result in significant activation in the liver orkidney. In other embodiments, some measurable extra-intestinal FXRactivation occurs, however the FXR activation is considerably greater inthe intestines than in other locations in the body, such as in the liveror kidney. In some embodiments, the FXR agonist is minimally absorbed.In some embodiments, the FXR agonist is directly administered to theintestines (such as to the distal ileum) of an individual in needthereof. In some embodiments, the FXR agonist is directly administeredto the colon or the rectum of an individual in need thereof. In someembodiments, the FXR agonist is administered orally, and less than 50%,less than 40%, less than 30%, less than 20%, less than 10%, less than9%, less than 8%, less than 7%, less than 6%, less than 5%, less than4%, less than 3%, less than 2%, or less than 1% of the FXR agonist issystemically absorbed.

In some examples, the subject to be treated is one who is diabetic (forexample has type II diabetes), is hyperglycemic, and/or is insulinresistant. In some examples, the subject is obese, for example has abody mass index (BMI) of 25 of higher, 30 or greater, 35 or greater, 40or greater, such as a BMI of 25 to 29, 30 to 34, or 35 to 40.

In some examples, the disclosed methods reduce weight gain in a subject(such as a human), such as diet-induced weight gain. In some examples,such methods reduce weight gain in the subject by at least 5%, at least10%, at least 15%, at least 20%, at least 30% or even at least 50% (suchas 5% to 50%, 5% to 25%, 10% to 20%, or 10% to 30%), for examplerelative to a subject not treated with the disclosed therapies.Similarly, in some examples, the disclosed methods reduce the BMI of asubject (such as a human). In some examples, such methods reduce the BMIof a subject by at least 5%, at least 10%, at least 15%, at least 20%,or at least 30% (such as 5% to 30%, 5% to 25%, 10% to 20%, or 10% to30%), for example relative to a subject not treated with the disclosedtherapies.

In some examples, the disclosed methods increase browning of whiteadipose tissue in a subject (such as a human). In some examples, suchmethods increase browning of white adipose tissue in the subject by atleast 5%, at least 10%, at least 15%, at least 20%, at least 30% or evenat least 50% (such as 5% to 50%, 5% to 25%, 10% to 20%, or 10% to 30%),for example relative to a subject not treated with the disclosedtherapies.

In some embodiments, the method reduces or prevents diet-induced weightgain, for example in a mammalian subject, such as a human. In someembodiments, the one or more FXR agonists are administered to an obesesubject whose obesity is diet-related (i.e., diet-induced obesity). Inother embodiments, the one or more FXR agonists can be administered toan obese subject whose obesity is not diet-related (such as anindividual with familial/genetic obesity or obesity resulting frommedication use). In other embodiments, the one or more FXR agonists canbe administered to a subject who is overweight (but not obese) or asubject that is neither overweight nor obese. Thus, in some embodiments,the one or more FXR agonists can be used to prevent obesity fromdeveloping. In some embodiments, the targeting of the therapy to theintestines reduces the chance of side effects which can result fromsystemic action, thus improving the safety profile of the therapy.

In some embodiments, the one or more FXR agonists are administered to anobese or non-obese subject for a metabolic disorder or condition otherthan obesity or weight gain. In certain embodiments, the metabolicdisorder is insulin resistance, including non-insulin-dependent diabetesmellitus (NIDDM) (i.e., type II diabetes). The administration of the oneor more FXR agonists can result in increased insulin sensitivity toinsulin in the liver, leading to increased uptake of glucose intohepatic cells. In certain embodiments, the metabolic disorder isdyslipidemia, including hyperlipidemia (elevated LDL, VLDL ortriglycerides) or low HDL levels. Thus, in certain embodiments,administration of one or more FXR agonists can result in improvedglucose and/or lipid homeostasis in the subject. In some embodiments,administration of the one or more FXR agonists results in a decrease inthe amount of hepatic triglycerides, serum lipids and/or triglyceridesin the subject. Thus, in some examples, the disclosed methods decreasethe amount of serum lipids and/or triglycerides in a subject (such as ahuman). In some examples, such methods decrease serum lipids and/ortriglycerides in the subject by at least 5%, at least 10%, at least 15%,at least 20%, at least 30%, at least 50% or even at least 75% (such as5% to 50%, 5% to 25%, 10% to 20%, 10% to 70%, or 10% to 30%), forexample relative to levels observed in a subject not treated with thedisclosed therapies. In some examples, such methods decrease hepatictriglycerides in the subject by at least 5%, at least 10%, at least 15%,at least 20%, at least 30%, at least 50% or even at least 75% (such as5% to 50%, 5% to 25%, 10% to 20%, 10% to 70%, or 10% to 30%), forexample relative to levels observed in a subject not treated with thedisclosed therapies. In some examples, the disclosed methods increaseinsulin sensitivity to insulin in the liver of a subject (such as ahuman) In some examples, such methods increase insulin sensitivity toinsulin in the liver of the subject by at least 5%, at least 10%, atleast 15%, at least 20%, at least 30% or even at least 50% (such as 5%to 50%, 5% to 25%, 10% to 20%, or 10% to 30%), for example relative to asubject not treated with the disclosed therapies.

In some embodiments, administration of the one or more FXR agonistsresults in no substantial change in food intake and/or fat consumptionin the subject. In other embodiments, food intake and/or fat consumptionis reduced minimally, such as by less than 15%, less than 10%, or lessthan 5%. In some embodiments, no substantial change in appetite in thesubject results. In other embodiments, reduction in appetite is minimalas reported by the subject.

In some embodiments, administration of the one or more FXR agonistsresults in an increase in the metabolic rate in the subject. Thus, insome examples, the disclosed methods increase the metabolic rate in asubject (such as a human) In some examples, such methods increase themetabolic rate in the subject by at least 5%, at least 10%, at least15%, at least 20%, at least 30%, at least 50% or even at least 75% (suchas 5% to 50%, 5% to 25%, 10% to 20%, 10% to 70%, or 10% to 30%), forexample relative to a subject not treated with the disclosed therapies.

In some embodiments, this increase in metabolism results from enhancedoxidative phosphorylation in the subject, which in turn can lead toincreased energy expenditure in tissues (such as BAT). Thus, in someexamples, the disclosed methods increase BAT activity in a subject (suchas a human). In some examples, such methods increase BAT activity in asubject by at least 5%, at least 10%, at least 15%, at least 20%, atleast 30%, at least 50% or even at least 75% (such as 5% to 50%, 5% to25%, 10% to 20%, 10% to 70%, or 10% to 30%), for example relative to asubject not treated with the disclosed therapies.

In some embodiments, administration of the one or more FXR agonistsresults in a decrease in the amount of serum insulin in the subject.Thus, in some examples, the disclosed methods decrease the amount ofserum insulin in a subject (such as a human) In some examples, suchmethods decrease serum insulin in the subject by at least 5%, at least10%, at least 15%, at least 20%, at least 30%, at least 50% or even atleast 75% (such as 5% to 50%, 5% to 25%, 10% to 20%, 10% to 70%, or 10%to 30%), for example relative to levels observed in a subject nottreated with the disclosed therapies.

In some embodiments, administration of the one or more FXR agonistsresults in a decrease in the amount of serum glucose in the subject.Thus, in some examples, the disclosed methods decrease the amount ofserum glucose in a subject (such as a human). In some examples, suchmethods decrease serum glucose in the subject by at least 5%, at least10%, at least 15%, at least 20%, at least 30%, at least 50% or even atleast 75% (such as 5% to 50%, 5% to 25%, 10% to 20%, 10% to 70%, or 10%to 30%), for example relative to levels observed in a subject nottreated with the disclosed therapies.Embodiments of a method areprovided for lowering elevations in blood glucose resulting from foodintake in a subject. Thus, in some examples, such methods decrease bloodglucose in a subject by at least 5%, at least 10%, at least 15%, atleast 20%, at least 30%, at least 50% or even at least 75% (such as 5%to 50%, 5% to 25%, 10% to 20%, 10% to 70%, or 10% to 30%), for examplerelative to a subject not treated with the disclosed therapies. Suchmethods can include orally administering to the subject atherapeutically effective amount of one of the disclosed minimallyabsorbed FXR agonists. In some embodiments, a method for loweringelevated body weight in a subject is provided, wherein the methodincludes orally administering to said subject a therapeuticallyeffective amount of one of the disclosed minimally absorbed FXRagonists. Thus, in some examples, such methods decrease the body weightof a subject by at least 5%, at least 10%, at least 15%, at least 20%,at least 30%, or at least 50% (such as 5% to 50%, 5% to 25%, 5% to 20%,10% to 20%, 10% to 70%, or 10% to 30%), for example relative to asubject not treated with the disclosed therapies. In some embodiments,the elevated body weight and/or elevated glucose levels resulted from aparticular pattern of food intake, such as a high fat diet and/or a highcalorie diet.

In some embodiments, the one or more FXR agonists are co-administeredwith one or more additional compounds or therapies, for treatment orprevention of a metabolic disorder. For example, the one or more FXRagonists can be administered with an insulin sensitizing drug, aninsulin secretagogue, an alpha-glucosidase inhibitor, a GLP agonist, aDPP-4 inhibitor (such as sitagliptin, vildagliptin, saxagliptin,linagliptin, anaglptin, teneligliptin, alogliptin, gemiglptin, ordutoglpitin), a catecholamine (such as epinephrine, norepinephrine, ordopamine), peroxisome proliferator-activated receptor (PPAR)-gammaagonist (e.g., a thiazolidinedione (TZD) [such as ioglitazone,rosiglitazone, rivoglitazone, or troglitazone], aleglitazar,farglitazar, muraglitazar, or tesaglitazar), or a combination thereof.Likewise, the one or more FXR agonists can be administered with astatin, HMG-CoA reductase inhibitor, fish oil, fibrate, niacin or othertreatment for dyslipidemia. In some embodiments, provided herein is amethod for treating a metabolic disorder in a subject, such as loweringelevated body weight and/or lowering elevated blood glucose from foodintake, comprising orally co-administering to said subject atherapeutically effective amount of a disclosed minimally absorbed FXRagonist and retinoic acid. 9 cis-retinoic acid is the ligand forretinoic acid receptor (RXR), the heterodimeric partner of FXR. In oneexample, nicotinamide ribonucleoside and/or analogs of nicotinamideribonucleoside that promotes NAD+ production of which is a substrate formany enzymatic reactions such as p450s which are a target of FXR (e.g.,see Yang et al., J. Med. Chem. 50:6458-61, 2007), are also administered.

Glucagon-like peptide-1 (GLP-1) is an incretin derived from thetranscription product of the proglucagon gene. The major source of GLP-1in the body is the intestinal L cell that secretes GLP-1 as a guthormone. The biologically active forms of GLP-1 include GLP-1-(7-37) andGLP-1-(7-36)NH₂ (HAEGTFTSDVSSYLEGQAAKEFIAWLVKGR; SEQ ID NO: 1), whichresult from selective cleavage of the proglucagon molecule. GLP-2 is a33 amino acid peptide (HADGSFSDEMNTILDNLAARDFINWLIQTKITD; SEQ ID NO: 2)in humans. GLP-2 is created by specific post-translational proteolyticcleavage of proglucagon in a process that also liberates GLP-1. GLPagonists are a class of drugs (“incretin mimetics”) that can be used totreat type 2 diabetes. Examples include, but are not limited to:exenatide (Byetta/Bydureon), liraglutide (Victoza), lixisenatide(Lyxumia), and albiglutide (Tanzeum). In certain embodiments, the FXRagonist enhances the secretion of glucagon-like peptide-1 (GLP-1) and/orglucagon-like peptide-2 (GLP-2). In some embodiments, the FXR agonistenhances the secretion of a pancreatic polypeptide-fold such as peptideYY (PYY). In certain embodiments, the FXR agonist enhances the activityof FGF15 or FGF19. In certain embodiments, the FXR agonist enhancessecretion of an enteroendocrine peptide and/or is administered incombination with an agent that enhances secretion or activity of anenteroendocrine peptide. Thus, in some examples, the disclosed methodsincrease the secretion of one or more of GLP-1, GLP-2, and PYY in asubject (such as a human). In some examples, such methods increase thesecretion of one or more of GLP-1, GLP-2, and PYY in the subject by atleast 5%, at least 10%, at least 15%, at least 20%, at least 30%, atleast 50% or even at least 75% (such as 5% to 50%, 5% to 25%, 10% to20%, 10% to 70%, or 10% to 30%), for example relative to a subject nottreated with the disclosed therapies. Furthermore, in some examples, thedisclosed methods increase the secretion of one or more of GLP-1, GLP-2,and PYY in a subject (such as a human). In some examples, such methodsincrease the activity of one or more of FGF15 and FGF19 in the subjectby at least 5%, at least 10%, at least 15%, at least 20%, at least 30%,at least 50% or even at least 75% (such as 5% to 50%, 5% to 25%, 10% to20%, 10% to 70%, or 10% to 30%), for example relative to a subject nottreated with the disclosed therapies.

The gut-biased FXR agonists disclosed herein can have profound metabolicbenefits with respect to obesity. The gut-biased FXR agonists canprotect against diet-induced weight gain by, for example, promoting theexpression of genes involved in thermogenesis, mitochondrial biogenesis,and/or fatty acid oxidation. In some embodiments, linked to theunexpected browning of white adipose, the disclosed gut-biased FXRagonists can lower inflammatory cytokine levels while up-regulatingβ-adrenergic signaling. These changes can be mediated, at least in part,by a change in bile acid levels and composition. In various embodiments,a prandial activation of intestinal FXR is triggered by administering toa subject one of the FXR agonists disclosed herein, such as syntheticFXR agonist fexaramine (Fex). The intestinal-specific FXR activationdisclosed herein can be utilized to enhance glucose tolerance and lowerhepatic glucose production. Thus, in some examples, such methodsdecrease hepatic glucose production in a subject by at least 5%, atleast 10%, at least 15%, at least 20%, at least 30%, at least 50% oreven at least 75% (such as 5% to 50%, 5% to 25%, 10% to 20%, 10% to 70%,or 10% to 30%), for example relative to a subject not treated with thedisclosed therapies. These physiologic changes can result in hepaticinsulin sensitization and/or BAT activation—properties not previouslyassociated with FXR agonists.

In contrast to the effects of system-wide drugs (including systemic FXRagonists), selective activation of intestinal FXR as disclosed hereincan mimic the restricted bile acid response linked to feeding. Thegut-specific FXR agonists disclosed herein robustly induce enteralFGF15, leading to alterations in bile acid composition withoutactivating hepatic FXR target genes. Unlike systemic drugs, these FXRagonists can protect against diet-induced weight gain, reduce body-wideinflammation, enhance thermogenesis, promote browning of white adiposetissue, promote activation of BAT, and suppress hepatic glucoseproduction.

In some embodiments, the initial event triggering systemic metabolicactivation is coordinated by FGF15 (the mouse equivalent of human FGF19)or FGF19. In an embodiment, administration of the FXR agonist results inactivation of FGF15 or FGF19 (such as an increase in FGF15 or FGF19activity of at least 25%, at least 50%, at least 75%, at least 90%, orat least 95%, relative to no treatment with an FXR agonist disclosedherein), which in turn can regulate energy expenditure, such as byincreasing metabolic rate, improving glucose homeostasis (such as byimproving insulin sensitivity), and/or improving lipid homeostasiswithout requiring significant changes in food intake. The absence of arequired or resulting change in food intake can be expected to increaseeffectiveness, as failure of appetite control is a major reason forweight gain and difficulty in losing weight. Thus, systemic increases inenergy expenditure, as seen in Fex-treated mice, can form the basis foran obesity treatment.

In some embodiments, treatment with one or more of the disclosed FXRagonists can produce a change in the bile acid pool, such as a dramaticincrease in the level of deoxycholic acid (such as an increase of atleast 25%, at least 50%, at least 75%, at least 90%, or at least 100%,relative to no treatment with an FXR agonist disclosed herein), a potentligand for the G protein-coupled bile acid receptor TGR5. Fex treatmentwas observed to induce D102, a downstream target of TGR5, in brownadipose tissue (BAT), thus implicating this additional pathway in theobserved increase in energy expenditure. Furthermore, the coordinate“browning” of white adipose tissue provides an independent yetcomplementary contribution to increased thermogenic capacity.

Thus, a new therapeutic avenue exists to manipulate energy expenditurewithout appetite changes through intestinally-biased activation of thenuclear receptor FXR. Furthermore, gut-restricted FXR agonists such asFex can offer improved safety profiles with limited circulation in theserum, thus reducing the risks of off-target effects and toxicity. Theremarkable metabolic improvements achieved with Fex treatment provide anew role for intestinal targeting in the control of metabolic disease.

B. Treatment or Prevention of Inflammation

Also provided herein are embodiments of a method for treating orpreventing an inflammatory intestinal condition. Certain disclosedembodiments can include administering a therapeutically effective amountof one or more FXR agonists to an individual in need thereof, such asone or more of the novel FXR agonists disclosed herein (such as 1, 2, 3,4 or 5 such agonists).

Thus, in some examples, the disclosed embodiments reduce inflammation ina subject (such as a human), such as inflammation in the intestine. Insome examples, disclosed embodiments reduce inflammation (such asintestinal inflammation) in the subject by at least 5%, at least 10%, atleast 15%, at least 20%, at least 30% or even at least 50% (such as 5%to 50%, 5% to 25%, 10% to 20%, or 10% to 30%), for example relative to asubject not treated with the disclosed therapies.

In various embodiments, the inflammatory condition can be necrotizingenterocolitis (NEC), gastritis, ulcerative colitis, inflammatory boweldisease, irritable bowel syndrome, pseudomembranous colitis,gastroenteritis, radiation induced enteritis, chemotherapy inducedenteritis, gastro-esophageal reflux disease (GERD), peptic ulcer,non-ulcer dyspepsia (NUD), celiac disease, intestinal celiac disease,gastrointestinal complications following bariatric surgery, gastriccarcinogenesis, or gastric carcinogenesis following gastric or bowelresection. In some embodiments, the inflammatory condition is NEC andthe subject is a newborn or prematurely born infant. In someembodiments, the subject is enterally-fed infant or formula-fed infant.

In some embodiments, the one or more FXR agonists are co-administeredwith one or more additional compounds or therapies, for treatment orprevention of an inflammatory intestinal condition. In some embodiments,the one or more FXR agonists are co-administered with an oralcorticosteroid and/or other anti-inflammatory or immuno-modulatorytherapy. In some embodiments, the FXR agonist can be administered to thesubject in conjunction with one or more antibiotics (e.g.,metronidazole, vancomycin, and/or fidaxomicin) to treat or prevent theinflammatory condition. In some embodiments, the FXR agonist can beadministered to the subject in conjunction with or following antibiotictherapy to treat or prevent pseudomembranous colitis associated withbacterial overgrowth (such as C. dificile overgrowth) in the subject. Insome embodiments, the FXR agonist can be administered to the subject inconjunction with metronidazole or other indicated therapy to treatinflammation associated with bacterial overgrowth in an intestinal area.In some embodiments, the FXR agonist can be administered to the subjectin conjunction with the ingestion of foods or other substances predictedto induce inflammation in the gastro-intestinal system of the subject(such as in a subject with celiac disease). In one example, nicotinamideribonucleoside and/or analogs of nicotinamide ribonucleoside thatpromotes NAD+ production of which is a substrate for many enzymaticreactions such as p450s which are a target of FXR (e.g., see Yang etal., J. Med. Chem. 50:6458-61, 2007), are also administered.

C. Prevention and/or Treatment of Cell Proliferation Diseases

Disclosed herein are embodiments of a method for preventing and/ortreating cell proliferation diseases, such as certain types of cancer.Certain disclosed embodiments can include administering atherapeutically effective amount of one or more FXR agonists to anindividual in need thereof, such as one or more of the novel FXRagonists disclosed herein (such as 1, 2, 3, 4 or 5 such agonists).

In some embodiments, the compounds disclosed herein may be used in theprevention or treatment of adenocarcinomas, i.e. carcinoma derived fromglandular tissue or in which the tumor cells form recognizable glandularstructures. Adenocarcinomas can be classified according to thepredominant pattern of cell arrangement, as papillary, alveolar, etc.,or according to a particular product of the cells, as mucinousadenocarcinoma. Adenocarcinomas arise in several tissues, including thecolon, kidney, breast, cervix, esophagus, gastric, pancreas, prostateand lung.

In certain embodiments, the compounds disclosed herein may be used inthe prevention or treatment of a cancer of the intestine, such as coloncancer, i.e. cancer that forms in the tissues of the colon (the longestpart of the large intestine), or a cancer of another part of theintestine, such as the jejunum, and/or ileum. Colon cancer is alsoreferred to as “colorectal cancer.” Most colon cancers areadenocarcinomas (cancers that begin in cells that may line internalorgans and have gland-like properties). Cancer progression ischaracterized by stages, or the extent of cancer in the body. Staging isusually based on the size of the tumor, whether lymph nodes containcancer, and whether the cancer has spread from the original site toother parts of the body. Stages of colon cancer include stage I, stageII, stage III and stage IV. In some embodiments herein, the colonadenocarcinoma is from any stage. In other embodiments, the colonadenocarcinoma is a stage I cancer, a stage II cancer or a stage IIIcancer.

Thus, in some examples, the disclosed embodiments reduce tumor burden ina subject (such as a human). In some examples, disclosed embodimentsreduce tumor burden (such as colon tumor burden) in the subject by atleast 5%, at least 10%, at least 15%, at least 20%, at least 30% or evenat least 50% (such as 5% to 50%, 5% to 25%, 10% to 20%, or 10% to 30%),for example relative to a subject not treated with the disclosedtherapies.

Thus, in some examples, the disclosed embodiments reduce tumor sizeand/or volume in a subject (such as a human) In some examples, disclosedembodiments reduce tumor size and/or volume (such as a colon tumor) inthe subject by at least 5%, at least 10%, at least 15%, at least 20%, atleast 30% or even at least 50% (such as 5% to 50%, 5% to 25%, 10% to20%, or 10% to 30%), for example relative to a subject not treated withthe disclosed therapies.

Thus, in some examples, the disclosed embodiments reduce effects ofcachexia due to a tumor in a subject (such as a human). In someexamples, disclosed embodiments reduce effects of cachexia (such as dueto a colon tumor) in the subject by at least 5%, at least 10%, at least15%, at least 20%, at least 30% or even at least 50% (such as 5% to 50%,5% to 25%, 10% to 20%, or 10% to 30%), for example relative to a subjectnot treated with the disclosed therapies.Thus, in some examples, thedisclosed embodiments increase survival rates of a subject (such as ahuman) with a tumor. In some examples, disclosed embodiments increasesurvival rates of a subject (such as a human) with a tumor (such as acolon cancer) in the subject by at least 5%, at least 10%, at least 15%,at least 20%, at least 30% or even at least 50% (such as 5% to 50%, 5%to 25%, 10% to 20%, or 10% to 30%), for example relative to a subjectnot treated with the disclosed therapies.

In some embodiments, the compounds disclosed herein may be administeredin combination with one or more additional anticancer therapies (such asa biologic [e. g., antibody, for example bevacizumab, cetuximab, orpanitumumab], chemotherapeutic, or radiologic, for example FOLFOX,FOLFIRI, CapeOX, 5-FU, leucovorin, regorafenib, irinotecan, andoxaliplatin), to prevent or treat a cell proliferation disease. In oneexample, nicotinamide ribonucleoside and/or analogs of nicotinamideribonucleoside that promotes NAD+ production of which is a substrate formany enzymatic reactions such as p450s which are a target of FXR (e.g.,see Yang et al., J. Med. Chem. 50:6458-61, 2007), are also administered.

D. Prevention and/or Treatment of Alcoholic and Non-Alcoholic LiverDisease

Disclosed herein are embodiments of a method for preventing and/ortreating alcoholic or non-alcoholic liver diseases, such as steatosis,cirrhosis, alcoholic hepatitis, NASH and NAFLD. In some embodiments, thecompounds disclosed herein may be used in the prevention or treatment ofalcoholic liver diseases. Certain disclosed embodiments can includeadministering a therapeutically effective amount of one or more FXRagonists to an individual in need thereof, such as one or more of thenovel FXR agonists disclosed herein (such as 1, 2, 3, 4 or 5 suchagonists).

Thus, in some examples, the disclosed embodiments reduce fatty liver(steatosis) in a subject (such as a human). In some examples, disclosedembodiments reduce steatosis in the subject (such as in an alcoholic) byat least 5%, at least 10%, at least 15%, at least 20%, at least 30% oreven at least 50% (such as 5% to 50%, 5% to 25%, 10% to 20%, or 10% to30%), for example relative to a subject not treated with the disclosedtherapies.

Thus, in some examples, the disclosed embodiments reduce cirrhosis in asubject (such as a human). In some examples, disclosed embodimentsreduce cirrhosis in the subject (such as in an alcoholic) by at least5%, at least 10%, at least 15%, at least 20%, at least 30% or even atleast 50% (such as 5% to 50%, 5% to 25%, 10% to 20%, or 10% to 30%), forexample relative to a subject not treated with the disclosed therapies.Thus the disclosed embodiments can reduce liver inflammation and/orfibrosis, for example by at least 5%, at least 10%, at least 15%, atleast 20%, at least 30% or even at least 50% (such as 5% to 50%, 5% to25%, 10% to 20%, or 10% to 30%), for example relative to a subject nottreated with the disclosed therapies.

Thus, in some examples, the disclosed embodiments reduce alcoholichepatitis in a subject (such as a human). In some examples, disclosedembodiments reduce alcoholic hepatitis in the subject (such as in analcoholic) by at least 5%, at least 10%, at least 15%, at least 20%, atleast 30% or even at least 50% (such as 5% to 50%, 5% to 25%, 10% to20%, or 10% to 30%), for example relative to a subject not treated withthe disclosed therapies. Thus the disclosed embodiments can reduceinflammation of hepatocytes, for example by at least 5%, at least 10%,at least 15%, at least 20%, at least 30% or even at least 50% (such as5% to 50%, 5% to 25%, 10% to 20%, or 10% to 30%), for example relativeto a subject not treated with the disclosed therapies.

Thus, in some examples, the disclosed embodiments reduce liver enzymesin a subject (such as a human). In some examples, disclosed embodimentsreduce liver enzymes (e.g., serum ALT and/or AST levels) in the subject(such as in an alcoholic) by at least 5%, at least 10%, at least 15%, atleast 20%, at least 30% or even at least 50% (such as 5% to 50%, 5% to25%, 10% to 20%, or 10% to 30%), for example relative to a subject nottreated with the disclosed therapies.

Thus, in some examples, the disclosed embodiments reduce livertriglycerides in a subject (such as a human). In some examples,disclosed embodiments reduce liver triglycerides in the subject (such asin an alcoholic) by at least 5%, at least 10%, at least 15%, at least20%, at least 30% or even at least 50% (such as 5% to 50%, 5% to 25%,10% to 20%, or 10% to 30%), for example relative to a subject nottreated with the disclosed therapies.

In some embodiments, the compounds disclosed herein may be administeredin combination with one or more additional therapies for treatingalcoholic or non-alcoholic liver disease (such as antioxidants,corticosteroids, and/or anti-TNF), to prevent or treat alcoholic ornon-alcoholic liver disease. In one example, nicotinamide ribonucleosideand/or analogs of nicotinamide ribonucleoside that promotes NAD+production of which is a substrate for many enzymatic reactions such asp450s which are a target of FXR (e.g., see Yang et al., J. Med. Chem.50:6458-61, 2007), are also administered.

E. Prevention and/or Treatment of Other Diseases

Disclosed herein are embodiments of a method for preventing and/ortreating cholestatic disorders, such primary biliary cirrhosis (PBC),primary sclerosing cholangitis (PSC), overlap syndrome (PBC plusautoimmune hepatitis),cholestasis resulting from a drug (e.g., one ormore of androgen, birth control pills, gold salts, nitrofurantoin,anabolic steroids, chlorpromazine, prochlorperazine, sulindac,cimetidine, estrogen, statins, and antibiotics such as TMP/SMX,flucoxacillin and erythromycin), drug-induced cholestatic hepatitis,total parenteral nutrition (TPN)-induced cholestasis, ICU/sepsis-relatedcholestasis, obstetric cholestasis, graft vs. host disease, prolongedcholestasis due to hepatitis A, B or C infection, cholestasis due tocystic fibrosis, alcoholic hepatitis, progressive familial intrahepaticcholestasis (PFIC) syndromes, Alagille syndrome, biliary atresia, or anycombination thereof. Certain disclosed embodiments can includeadministering a therapeutically effective amount of one or more FXRagonists to an individual in need thereof, such as one or more of thenovel FXR agonists disclosed herein (such as 1, 2, 3, 4 or 5 suchagonists). Thus, in some examples, the disclosed embodiments increasebile flow in a subject (such as a human) by at least 5%, at least 10%,at least 15%, at least 20%, at least 30%, at least 40%, at last 50%, atleast 60%, at least 70%, at least 80%, at least 90%, at least 100%, oreven at least 200% (such as 5% to 50%, 5% to 25%, 50% to 75%, or 75% to200%), for example relative to a subject not treated with the disclosedtherapies.

Disclosed herein are embodiments of a method for preventing and/ortreating an intestinal permeability condition, such as Crohn's disease,ulcerative colitis, infectious colitis, celiac disease, type 1 diabetes,inflammatory bowel disease, irritable bowel syndrome, or any combinationthereof. Certain disclosed embodiments can include administering atherapeutically effective amount of one or more FXR agonists to anindividual in need thereof, such as one or more of the novel FXRagonists disclosed herein (such as 1, 2, 3, 4 or 5 such agonists). Insome examples, disclosed embodiments reduce undesired intestinalpermeability in the subject by at least 5%, at least 10%, at least 15%,at least 20%, at least 30% or even at least 50% (such as 5% to 50%, 5%to 25%, 10% to 20%, or 10% to 30%), for example relative to a subjectnot treated with the disclosed therapies.

Disclosed herein are embodiments of a method for preventing and/ortreating a disorder that causes or results from an altered intestinalmicrobiome, such as celiac disease, the intestinal permeabilityconditions described herein, the intestinal inflammation disordersdescribed herein, alcoholic hepatitis, necrotizing enterocolitis,Crohn's disease, ulcerative colitis, intestinal lesions (such as thosein a cystic fibrosis patient), cirrhosis, or any combination thereof.Certain disclosed embodiments can include administering atherapeutically effective amount of one or more FXR agonists to anindividual in need thereof, such as one or more of the novel FXRagonists disclosed herein (such as 1, 2, 3, 4 or 5 such agonists). Insome examples, disclosed embodiments bring the intestinal microbiomecloser to normal levels in the subject, for example within 20%, with in10% or within 5% of normal for example relative to a subject not treatedwith the disclosed therapies.

Disclosed herein are embodiments of a method for treating an inbornerror of metabolism, such as cerebrotendinous xanthomatosis. Certaindisclosed embodiments can include administering a therapeuticallyeffective amount of one or more FXR agonists to an individual in needthereof, such as one or more of the novel FXR agonists disclosed herein(such as 1, 2, 3, 4 or 5 such agonists). In some examples, disclosedembodiments reduce plasma cholesterol levels in the subject by at least5%, at least 10%, at least 15%, at least 20%, at least 30% or even atleast 50% (such as 5% to 50%, 5% to 25%, 10% to 20%, or 10% to 30%), forexample relative to a subject not treated with the disclosed therapies.

Disclosed herein are embodiments of a method for treating a biledisorder, such as benign biliary stricture, malignant biliaryobstruction, bile acid diarrhea, or any combination thereof. Certaindisclosed embodiments can include administering a therapeuticallyeffective amount of one or more FXR agonists to an individual in needthereof, such as one or more of the novel FXR agonists disclosed herein(such as 1, 2, 3, 4 or 5 such agonists). In some examples, disclosedembodiments reduce production of bile acids in the subject by at least5%, at least 10%, at least 15%, at least 20%, at least 30% or even atleast 50% (such as 5% to 50%, 5% to 25%, 10% to 20%, or 10% to 30%), forexample relative to a subject not treated with the disclosed therapies.In some examples, disclosed embodiments increase intestinal absorptionof bile acids in the subject by at least 5%, at least 10%, at least 10%,at least 15%, at least 20%, at least 30%, at least 40%, at last 50%, atleast 60%, at least 70%, at least 80%, at least 90%, at least 100%, oreven at least 200% (such as 5% to 50%, 5% to 25%, 50% to 75%, or 75% to200%), for example relative to a subject not treated with the disclosedtherapies.

Disclosed herein are embodiments of a method for treating or preventinga malabsorption disorder (e.g., intestinal malabsorption), such as shortbowel syndrome (or symptoms arising from such, such as diarrhea,steatorhea, malnutrition, fatigue, vitamin deficiency), environmentalenteropathy, or tropical sprue. Certain disclosed embodiments caninclude administering a therapeutically effective amount of one or moreFXR agonists to an individual in need thereof, such as one or more ofthe novel FXR agonists disclosed herein (such as 1, 2, 3, 4 or 5 suchagonists). In some examples, disclosed embodiments increase bowelabsorption in the subject by at least 5%, at least 10%, at least 10%, atleast 15%, at least 20%, at least 30%, at least 40%, at last 50%, atleast 60%, at least 70%, at least 80%, at least 90%, at least 100%, oreven at least 200% (such as 5% to 50%, 5% to 25%, 50% to 75%, or 75% to200%), for example relative to a subject not treated with the disclosedtherapies.

F. Administration

The particular mode of administration and the dosage regimen will beselected by the attending clinician, taking into account the particularsof the case (e.g. the subject, the disease, the disease state involved,the particular treatment, and whether the treatment is prophylactic).Treatment can involve daily or multi-daily or less than daily (such asweekly or monthly etc.) doses over a period of a few days to months, oreven years. For example, a therapeutically effective amount of one ormore compounds disclosed herein can be administered in a single dose,twice daily, weekly, or in several doses, for example daily, or during acourse of treatment. In a particular non-limiting example, treatmentinvolves once daily dose or twice daily dose.

In some embodiments, the FXR agonist(s) is administered orally. In someembodiments, the FXR agonist is administered as an ileal-pH sensitiverelease formulation that delivers the FXR agonist to the intestines,such as to the ileum of an individual. In some embodiments, the FXRagonist is administered as an enterically coated formulation. In someembodiments, oral delivery of an FXR agonist provided herein can includeformulations, as are well known in the art, to provide prolonged orsustained delivery of the drug to the gastrointestinal tract by anynumber of mechanisms. These include, but are not limited to, pHsensitive release from the dosage form based on the changing pH of thesmall intestine, slow erosion of a tablet or capsule, retention in thestomach based on the physical properties of the formulation, bioadhesionof the dosage form to the mucosal lining of the intestinal tract, orenzymatic release of the active drug from the dosage form. The intendedeffect is to extend the time period over which the active drug moleculeis delivered to the site of action (e.g., the intestines) bymanipulation of the dosage form. Thus, enteric-coated and enteric-coatedcontrolled release formulations are within the scope of the presentdisclosure. Suitable enteric coatings include cellulose acetatephthalate, polyvinylacetate phthalate, hydroxypropylmethylcellulosephthalate and anionic polymers of methacrylic acid and methacrylic acidmethyl ester.

In some embodiments, the FXR agonist is administered before ingestion offood, such as at least 10 minutes, at least 15 minutes, at least 20minutes, or at least 30 minutes before ingestion of food (such as 10-60minutes or 10-30 minutes before ingesting food). In some embodiments ofthe methods described herein, the FXR agonist is administered less thanabout 60 minutes before ingestion of food. In some embodiments of themethods described above, the FXR agonist is administered less than about30 minutes before ingestion of food. In some embodiments of the methodsdescribed herein, the FXR agonist is administered after ingestion offood. In some embodiments, the methods further comprise administrationof a DPP-IV inhibitor, a TGR5 agonist, a biguanide, an incretin mimetic,or GLP-1 or an analog thereof. In some embodiments, the methods furthercomprise administration of a steroid or other anti-inflammatory compoundwhich may have an effect in the gut. In some embodiments, the methodsfurther include co-administration of an antibiotic therapy, and the FXRagonist treats or prevents inflammation, such as inflammation associatedwith antibiotic-induced colitis.

The composition administered can include at least one of a spreadingagent or a wetting agent. In some embodiments, the absorption inhibitoris a mucoadhesive agent (e.g., a mucoadhesive polymer). In someembodiments, the mucoadhesive agent is selected from methyl cellulose,polycarbophil, polyvinylpyrrolidone, sodium carboxymethyl cellulose, anda combination thereof. In some embodiments, a pharmaceutical compositionadministered further includes an enteroendocrine peptide and/or an agentthat enhances secretion or activity of an enteroendocrine peptide.

The pharmaceutical compositions that comprise one or more compoundsdisclosed herein can be formulated in unit dosage form, suitable forindividual administration of precise dosages. In one non-limitingexample, a unit dosage contains from about 1 mg to about 50 g of one ormore compounds disclosed herein, such as about 10 mg to about 10 g,about 100 mg to about 10 g, about 100 mg to about 1 g, about 500 mg toabout 5 g, or about 500 mg to about 1 g. In other examples, atherapeutically effective amount of one or more compounds disclosedherein is from about 0.01 mg/kg to about 500 mg/kg, for example, about0.5 mg/kg to about 500 mg/kg, about 5 mg/kg to about 250 mg/kg, or about50 mg/kg to about 100 mg/kg. In other examples, a therapeuticallyeffective amount of one or more compounds disclosed herein is from about50 mg/kg to about 250 mg/kg, for example about 100 mg/kg.

VII. WORKING EXAMPLES Example 1 Activity of Orally-administeredFexaramine is Restricted to the Intestine

Upon exploration of the in vivo effects of fexaramine (Fex)administration, it was discovered that due to ineffectual absorption,oral (PO) and intraperitoneal (IP) drug delivery produced very differenteffects (FIGS. 1D and 1E). While robust induction of the FXR target geneSHP was seen throughout the intestine with both acute PO and IP Fextreatment (100 mg/kg for five days), induction of SHP was only seen inliver and kidney after IP treatment (FIG. 1A). Consistent with thisnotion, PO Fex treatment induced multiple FXR target genes in theintestine including IBABP, OSTα and FGF15, but failed to affect theexpression of these genes in liver or kidney (FIGS. 1B, 1C and 1F).Quantification of serum Fex levels revealed an order of magnitude lowerdrug levels after acute PO- compared to IP-treatment (−10% of IP levels)(FIGS. 1D and 1E). Notably, the serum levels of Fex after POadministration were below the 25 nM EC₅₀ of Fex, consistent with thelack of target gene activation in the kidney and liver.

Example 2 Fexaramine Prevents Diet-induced Obesity Weight Gain

To investigate the physiological effects of intestinal FXR activation byfexaramine, mice were subjected to chronic fexaramine (100 mg/kg Fex) POtreatment for 5 weeks. Chronically treated chow-fed mice wereindistinguishable from vehicle-treated mice in terms of weight gain,basal metabolic activity and glucose tolerance (FIGS. 3A-3D).

The physiological effects of fexaramine in established obesity(diet-induced obesity, DIO) models were evaluated. C57BL/6J mice werefed a diet of 60% fat for 14 weeks and then treated PO with vehicle orfexaramine (100 mg/kg) for 5 weeks. Surprisingly, chronic fexaramineoral administration prevented weight gain in DIO mice (FIG. 2A).Prevention of weight gain by fexaramine occurred in a dose-dependentmanner (FIG. 4A) with no signs of intestinal toxicity (FIG. 4B). At thehighest dose weight gain was almost completely abrogated. The reductionin weight gain of Fex-treated mice was largely attributed to reducedoverall fat mass (as analyzed by MRI), with significant reductions inwet weights of both subcutaneous (inguinal) and visceral (gonadal andmesenteric) adipose depots (FIGS. 2B and 2C). Consistent with reducedadiposity, Fex-treated mice showed significant improvements in theirendocrine and metabolic profiles including reduced glucose, insulin,leptin, cholesterol, and resistin levels (FIGS. 2D and 4D).

Obesity and its metabolic complications are associated with chroniclow-grade inflammation, reflected by elevated serum levels ofinflammatory cytokines. Serum levels of inflammatory cytokines TNFα,IL-1α, IL-1β, IL-17 and MCP-1 were markedly decreased by fexaramine(FIG. 2E) (such as reductions of at least 50%, at least 75%, at least80%, or even at least 90%), indicating that fexaramine-induced weightgain resistance reduced systemic inflammation.

The reduction in fasting insulin levels also suggested improved glucosetolerance and insulin sensitivity in fexaramine-treated DIO mice.Therefore, glucose tolerance tests (GTTs) and insulin tolerance tests(ITTs) were performed to determine if glucose homeostasis was improvedin fexaramine-treated DIO mice. Fex treatment induced dose-dependentimprovements in glucose tolerance and insulin sensitivity in DIO mice(measured by glucose and insulin tolerance tests) (FIGS. 2F and 2G and4C). In addition, while fexaramine improved glucose homeostasis in adose-dependent manner in DIO mice, there were no effects observed innormal chow-fed mice across a range of doses. Notably, these Fex-inducedchanges in gene expression and improvements in metabolic homeostasiswere abrogated in Fex-treated FXR null mice, establishing the FXRdependence of the observed effects (FIGS. 5A-5I).

Example 3 Fexaramine Enhances Energy Expenditure in Brown Adipose Tissue

As the differential weight effect was not attributable to difference infood intake between vehicle-treated control mice and Fex-treated mice(FIG. 6A), the metabolic rates of weight-matched mice were compared.Fex-treated DIO mice had consistently higher oxygen consumption (VO₂)and exhaled more carbon dioxide (VCO₂) than vehicle-treated controls(FIGS. 6B-6C), but displayed similar respiratory exchange ratios,suggesting enhanced metabolism of both sugar and fat (FIG. 6M). Based onambulatory counts, Fex-treated mice were more active than control mice,which can be a result of lower body weights supporting increased energyexpenditure in treated mice (FIG. 6D).

Consistent with increased energy expenditure, Fex treatment increasedthe core body temperature approximately 1.5° C. (FIG. 6E). In addition,the prominent accumulation of lipid vesicles in brown adipose tissue(BAT) of vehicle-treated DIO mice was markedly reduced in Fex-treatedmice (FIG. 6F). Gene expression analysis confirmed the induction ofERRγ, PGC-1α, and PGC-10, as well as a number of their target genesinvolved in thermogenesis, mitochondrial biogenesis, and fatty acidoxidation in BAT (FIG. 6G). Moreover, Fex treatment increased thephosphorylation level of p38 (FIG. 6H and 6I), previously shown tostabilize PGC-1α, a key coactivator of the thermogenic transcriptionalprogram in BAT. A comparison of the transcriptional changes induced byFex in inguinal, gonadal and brown adipose depots revealed coordinatedchanges that selectively enhance OXPHOS activity only in BAT, indicatingthat BAT is a key contributor to the increased energy expenditure andthermogenesis (FIG. 6J). Consistent with this conclusion, KEGG pathwayanalysis of Fex-induced transcriptional changes from RNA-sequenceanalysis in BAT identified oxidative phosphorylation as significantlychanged (Table 1), and increased PKA activity was seen in Fex-treatedmice (FIG. 6L).

TABLE 1 KEGG pathway Term p-value Oxidative phosphorylation 8.12E−07Chemokine signaling pathway 2.21E−03 Cytokine-cytokine receptorinteraction 4.40E−03 Biosynthesis of unsaturated fatty acids 7.04E−03PPAR signaling pathway 7.53E−03

Furthermore, serum lactate levels were significantly reduced inFex-treated DIO mice, suggesting that body-wide energy metabolism isshifted towards a more oxidative state (FIG. 6N). Thus, the markedreduction in lipids, increased PKA activity and p38 phosphorylation, andincreased core body temperature indicate a coordinated activation ofthermogenesis in BAT in Fex-treated DIO mice.

Example 4 Fexaramine Induces FGF15 and Alters Bile Acid Composition

RNA-Seq of intestinal tissues was used to explore the mechanisms throughwhich Fex might contribute to systemic changes in energy expenditure andmetabolic rate. Mice were fed on HFD for 14 weeks, and then subjected todaily oral injection of vehicle or fexaramine (100 mg/kg) for 5 weekswith HFD. KEGG pathway analysis revealed the induction of multiplecellular metabolic pathways including PPAR and adipocytokine signalingin both ileum and colon (Tables 2 and 3).

TABLE 2 KEGG pathway (ileum) KEGG pathway Term p-value PPAR signalingpathway 1.86E−05 Adipocytokine signaling pathway 2.91E−03 Retinolmetabolism 3.03E−03 Drug metabolism 4.01E−03 Arachidonic acid metabolism5.33E−03

TABLE 3 KEGG pathway (colon) KEGG pathway Term p-value PPAR signalingpathway 3.52E−11 Adipocytokine signaling pathway 8.90E−03 Retinolmetabolism 7.06E−02

Overlap of Fex-induced expression changes with previously identifiedintestinal FXR binding sites identified a subset of genes as potentialdirect FXR target genes (FIG. 7A). Within this subset, FGF15(corresponds to FGF19 in humans) was found to be dramaticallyup-regulated by Fex. In addition to established FXR target genes such asLp1, other genes exhibiting regulation by FXR were identified includingPer1 (FIG. 7A).

As an intestinal endocrine hormone, FGF15 induction is of interest sinceit activates the thermogenic program in BAT, as well as negativelyregulate BA synthesis through suppression of hepatic CYP7A1, therate-limiting enzyme for BA synthesis. An increase in circulating FGF15accompanied the increase in mRNA expression in ileum (FIGS. 7B and 7C)(such as an increase of at least 100%, at least 125%, or at least 150%).Consistent with an increase in serum FGF15, hepatic CYP7A1 expressionwas significantly repressed at both the mRNA and protein level afterchronic Fex treatment, while the expression of CYP8B1 and CYP27A1(enzymes not regulated by FGF15) were not affected (FIG. 7D and FIG. 8).In addition, expression of established liver FXR target genes SHP andBSEP were not altered, further demonstrating the absence of hepatic FXRactivation after chronic Fex treatment (FIG. 7D) and indicating thatother pathways, such as FGF15, mediate changes in hepatic geneexpression.

Genetic activation of intestinal FXR has been previously shown to alterbile acid composition. This is relevant as dietary, microbial or hepaticstress can alter the pool and enhance the production of toxic andcholestatic BAs such as taurine-conjugated chenodeoxycholic acid(T-CDCA) and taurine-conjugated cholic acid (T-CA). Despite the apparentabsence of hepatic FXR activation, Fex treatment produced strikingchanges in the composition of the BA pool. In addition to reducing thebile acid pool size, Fex treatment changed the relative proportions ofcirculating bile acids, most notably decreasing the fraction oftaurocholic acid and increasing the fraction of the secondary bile acid,lithocholic acid (FIGS. 7E and 7F, Table 4). These changes are inkeeping with increased intestinal FXR activation, including the effectsof increased circulating FGF15 on bile acid synthesis in the liver.Indeed, decreased serum taurocholic acid has been previously reported inmice expressing a constitutively activated FXR transgene in intestine,as well as after injection of FGF19, the human analogue of FGF15 (Wu etal. PloS one 6, e17868, 2011). Furthermore, changes in bile acidsynthesis away from cholic acid towards chenodeoxycholic acid and itsderivatives, which includes lithocholic acid, were observed upon FGF19treatment, consistent with a reduction in hepatic CYP7A1 and an increasein CYP7B1 expression.

TABLE 4 Fexaramine alters the serum bile acid composition Bile AcidComposition (%) Vehicle Fexaramine CA 4.08 7.51 TCA 34.96 12.23 CDCA1.86 2.51 TCDCA 3.52 1.13 LCA 7.67 28.13 GLCA N.D. 0.51 DCA 6.03 7.67TDCA 1.42 1.02 HDCA 1.20 0.36 T-HDCA 0.99 N.D UDCA 0.01 0.05 T-UDCA 2.853.07 alpha MCA 0.33 N.D beta MCA 0.55 N.D T-beta MCA 31.78 29.16 omegaMCA 2.74 6.65Mice fed a HFD for 14 weeks were maintained on a HFD and treated withvehicle or fexaramine (100 mg/kg/day per os for 5 week). Serum bile acidcomposition was determined by mass spectrometry. N.D not determined.

FXR activation has been reported to enhance mucosal defense geneexpression and intestinal barrier function (Inagaki et al., Proc NatlAcad Sci U S A 103:3920-3925, 2006; Gadaleta., et al. Gut 60:463-472,2011). Consistent with these reports, mice showed reduced intestinalpermeability, as measured by FITC-dextran leakage into the serum, andincreased expression of mucosal defense genes Occludin and Muc2, afterchronic Fex-treatment (FIGS. 7G and 7H).

While Fex does not activate the G protein-coupled bile acid receptor,TGR5 (FIG. 9), the pronounced changes in BAs indicated that this pathwaymay contribute to the observed physiologic effects. Notably, treatmentof HFD-fed mice with the intestinally-restricted TGR5 agonist, L7550379,failed to induce metabolic changes, while treatment with the systemicTGR5 agonist, RO5527239 improved glucose homeostasis, as measured by GTTand insulin secretion (FIGS. 10A-10F). These results indicated that TGRSactivation outside of the intestine may contribute to the beneficialeffects of Fex treatment (FIGS. 10B, 10D, 10E and 10F).

To address this possibility, HFD-fed TGRS null mice were chronicallytreated with Fex (100 mg/kg/day PO for 5 weeks). As seen in wild typemice, Fex treatment induced multiple FXR target genes in the ileum ofTGRS null mice including FGF15, resulting in lowered serum BA levels(FIGS. 11A, 11B). In this TGRS null background, Fex treatment inducedmoderate improvements in fasting glucose levels and glucose tolerance(FIGS. 11C, 11D). In addition, somewhat blunted increases in core bodytemperature and metabolic rate, correlating with the induction ofthermogenic genes in BAT, were observed (FIGS. 11E-11H), indicating thatthese effects do not require TGRS activation. In contrast to wild typemice, no significant changes in weight gain or insulin sensitivity wereobserved in Fex treated TGRS null mice, and altered gene expressionpatterns were seen in the liver and muscle, indicating involvement ofthe TGRS pathway (FIGS. 11I-11N). In particular, the anti-lipogeniceffects of Fex in the liver appear to require TGR5 activation, as keyhepatic lipogenic genes and liver triglyceride content were not affectedby Fex treatment (FIGS. 11L, 11M).

Example 5 Fexaramine Induces Browning of White Adipose Tissue

During obesity, adipose tissue expands by hyperplastic and/orhypertrophic growth, is chronically inflamed, and produces inflammatorycytokines that ultimately contribute to systemic metabolicdysregulation. After chronic Fex-treatment, the cross-sectional area ofadipocytes in visceral depots including gonadal and mesenteric wasmarkedly reduced (FIG. 12A). Investigation of signaling pathwaysimplicated in diet-induced inflammation identified reduced levels ofIKK-ε and TANK-binding kinase 1 (TBK1) in Fex-treated DIO mice (FIGS.12B, 13). These noncanonical IκB kinases were recently shown to playcrucial roles in energy expenditure as a consequence of adipose tissueinflammation upon diet-induced obesity (Reilly et al., Nat Med19:313-321, 2013). In addition, activation of the mammalian target ofrapamycin complex1 (mTORC1) pathway, a key lipogenic pathway activatedby high fat diet (HFD), was reduced in Fex-treated gonadal WAT, asevidenced by reduced S6K phosphorylation (FIG. 12B). Consistent withreduced adiposity, expression of the inflammatory cytokines TNFα, MCP-1and IL-1α, as well as the macrophage marker F4/80, were reduced invisceral and brown adipose depots of Fex-treated mice (FIGS. 12C and14).

Brown adipose-driven adaptive thermogenesis is fueled by mitochondrialoxidation of free fatty acids (FFAs) released from triglyceride storesinto the circulation predominantly by the action of hormone-sensitivelipase (HSL). Low levels of HSL phosphorylation were seen in visceraland subcutaneous adipose depots from control mice, as expected, due todesensitization of the β-adrenergic pathway in WAT during obesity(Carmen & Victor, Cell Signal 18:401-408, 2006; Song et al. Nature468:933-9, 2010). In contrast, a pronounced increase in HSLphosphorylation and serum levels of free fatty acids (FIGS. 12D and12G), accompanied by increased serum catecholamine levels andβ3-adrenergic receptor expression (FIGS. 12C, 12E and 12F), was observedafter chronic Fex treatment. As β-adrenergic receptor activation hasbeen shown to induce “brown fat-like” cells in inguinal adipose tissue,and these cells have been associated with resistance to diet-inducedobesity and improved glucose metabolism (Tsukiyama-Kohara et al., NatMed 7:1128-1132, 2001; Fisher et al., Genes Dev 26:271-281, 2012; Hansenet al., Proc Natl Acad Sci U S A 101:4112-4117, 2004; Wang et al., MolCell Biol 28:2187 -2200, 2008), UCP-1 expression was examined ininguinal adipose tissue. Immunohistochemistry revealed a substantialincrease in the abundance of multi-locular, UCP1-expressing adipocytesin Fex-treated animals (FIG. 12H). Furthermore, Fex-treatment increasedthe expression of “brown fat-like” signature genes, as well as increasedrespiratory capacity in the stromal vascular fraction from inguinaladipose tissue (FIGS. 12I and 12J). These results indicate thatFexaramine, unlike systemic FXR ligands, induces a distinct coordinatedmetabolic response, enhancing (3adrenergic signaling to promotelipolysis, mobilizing fatty acids for oxidation in BAT and the“browning” of cells in white adipose tissue.

Example 6 Fexaramine Improves Insulin Sensitivity and Glucose Tolerance

To probe the mechanism through which chronic Fex treatment improvedglucose homeostasis, hyperinsulinemic-euglycemic clamp studies wereperformed. No differences in basal hepatic glucose production (HGP),glucose disposal rate (GDR), insulin-stimulated GDR (IS-GDR), free fattyacid (FFA) suppression, and fasting insulin levels were observed betweenweight-matched cohorts (generated by treating initially heavier mice(2-3 grams) with Fex (FIGS. 15A-15C, FIGS. 15I and 15K)). However,Fex-treated mice displayed a marked increase in insulin-mediatedsuppression of HGP compared to control DIO mice (FIG. 15D). Thus, whilethe attenuated weight gain can contribute to improved glucose clearancein Fex-treated mice, this improvement in hepatic glucose suppressionindicates enhanced liver insulin sensitivity after Fex treatment.

Liver insulin resistance has been linked to obesity-induced hepaticsteatosis (Cohen et al., Science 332:1519-1523, 2011). Histologicalexamination of liver tissue from Fex-treated DIO mice revealed areduction in lipid droplets compared to controls indicating ameliorationof hepatic steatosis (FIG. 15E). Consistent with this histology, amarked decrease in hepatic triglycerides (such as a reduction of atleast 10%, or at least 20%) and reduced hepatic expression ofgluconeogenic and lipogenic genes (such as a reduction of at least 20%,or at least 30%, or at least 50%) were seen after chronic Fex treatment(FIGS. 15F and 15G). Furthermore, decreased serum alanineaminotransferase (ALT) levels were measured in Fex-treated mice,indicating reduced HFD-induced liver damage (FIG. 5H). Thus, in DIO miceFex promotes hepatic insulin sensitization, reduced steatosis, improvedmetabolic markers, decreased ALT and enhanced BAT activity.

Example 7 FXR Activity Screen for Determining EC₅₀ Determination

Cell Culture and Transfection: CV-1 cells were grown in DMEM+10%charcoal stripped FCS. Cells were seeded into 384-well plates the daybefore transfection to give a confluency of 50-80% at transfection. Atotal of 0.8 grams DNA containing 0.32 micrograms pCMX-hFXRfl, 0.32micrograms pCMX-hRXRfl, 0.1 micrograms pCMX.beta.Gal, 0.08 microgramspGLFXRE reporter and 0.02 micrograms pCMX empty vector was transfectedper well using FuGene transfection reagent according to themanufacturer's instructions (Roche). Cells were allowed to expressprotein for 48 hours followed by addition of compound.

Plasmids: Human FXR full length and RXR full length was obtained fromRonald Evans' laboratory and PCR amplification of the hFXR cDNA and thehRXR cDNA was performed. The amplified cDNAs was cloned into the vectorpCMX generating the plasmids pCMX-hFXRfl and pCMX-hRXRfl. Ensuingfusions were verified by sequencing. The pCMXMH2004 luciferase reportercontains multiple copies of the GAL4 DNA response element under aminimal eukaryotic promoter (Hollenberg and Evans, 1988). pCMX.beta.Galwas generated in the Evans laboratory, Salk Institute.

Compounds: All compounds were dissolved in DMSO and diluted 1:1000 uponaddition to the cells. Compounds were tested in quadruple inconcentrations ranging from 0.001 to 100 μM. Cells were treated withcompound for 24 hours followed by luciferase assay. Each compound wastested in at least two separate experiments.

Luciferase assay: Medium including test compound was aspirated andwashed with PBS. 50 μL PBS including 1 mM Mg²⁺ and Ca²⁺ were then addedto each well. The luciferase assay was performed using the LucLite kitaccording to the manufacturer's instructions (Packard Instruments).Light emission was quantified by counting on a Perkin Elmer Envisionreader. To measure 3-galactosidase activity 25 μL supernatant from eachtransfection lysate was transferred to a new 384 microplate.Beta-galactosidase assays were performed in the microwell plates using akit from Promega and read in a Perkin Elmer Envision reader. Thebeta-galactosidase data were used to normalize (transfection efficiency,cell growth etc.) the luciferase data.

Statistical Methods: The activity of a compound is calculated as foldinduction compared to an untreated sample. For each compound theefficacy (maximal activity) is given as a relative activity compared toFexaramine, a FXR agonist. The EC₅₀ is the concentration giving 50% ofmaximal observed activity. EC₅₀ values were calculated via non-linearregression using GraphPad PRISM (GraphPad Software, San Diego, Calif.).The EC₅₀ values for exemplary compounds are given in Table 5.

TABLE 5 purity, yields and activity data of exemplary fexaramine analogsPurity 1st RND 2nd RND Code number ELSD/UV yield (mg) EC₅₀ EC₅₀ Fex EC₅₀NSSK00004 99.9 10.8 237 nM 266.2 nM 31.9/36 nM NSSK00017 96.2 5.8 756 nM500 nM 31.9/36 nM NSSK00035 96.2 8.6 3.3 μM 4.49 μM 31.9/36 nM NSSK0000598.7 9 1.82 μM 2.1 μM 31.9/36 nM NSSK00018 98.6 6.8 380.5 nM 415.2 nM31.9/36 nM NSSK00006 100.0 17 273.1 nM 242.8 nM 31.9/36 nM NSSK00019100.0 7.7 923 nM 554 nM 53.7/39 nM NSSK00036 99.7 4.3 18.6 uM 4.9 uM53.7/39 nM NSSK00008 99.1 10.9 2.0 uM 1.4 uM 53.7/39 nM NSSK00007 99.913.9 473 nM 169.4 nM 53.7/39 nM NSSK00020 98.3 6.1 743 nM 463.7 nM53.7/39 nM NSSK00009 97.2 4.7 3.3 uM 4.9 uM 53.7/39 nM NSSK00022 97.8 33.2 uM 2.7 uM 56/52.5 nM NSSK00037 98.2 2.8 1.9 mM 4.8 mM 56/52.5 nMNSSK00001 95.2 11.5 68.6 nM 55.8 nM 56/52.5 nM NSSK00002 97.4 8.3 96.8nM 65.9 nM 56/52.5 nM NSSK00033 100.0 10 169.5 nM 254.4 nM 56/52.5 nMNSSK00034 100.0 6.1 2.2 uM 2.68 uM 56/52.5 nM NSSK00012 99.7 7.7 1.2 uM1.2 uM 39.9/34 nM NSSK00011 100.0 18.5 288.7 nM 379 nM 39.9/34 nMNSSK00014 95.8 12.2 345.6 nM 475.1 nM 39.9/34 nM NSSK00013 99.9 9.6390.3 nM 429.8 nM 39.9/34 nM NSSK00016 99.9 15.5 284.9 nM 377.4 nM39.9/34 nM NSSK00026 98.1 12.3 587.2 nM 910.5 nM 39.9/34 nM NSSK0002594.4 3.1 150.4 nM 167.7 nM 36.4/33 nM NSSK00024 99.5 15.8 66.8 nM 58.5nM 36.4/33 nM (Salk 00024) NSSK00024 83 nM 63 nM (retested) NSSK0002796.8 19.4 48.9 nM 46.6 nM 36.4/33 nM (Salk 00027) NSSK00027 118 nM 63 nM(retested) NSSK00030 95.8 9.2 655.5 nM 375 nM 36.4/33 nM NSSK00029 99.617.9 605 nM 510 nM 36.4/33 nM NSSK00031 96.5 18.6 366 nM 249 nM 36.4/33nM NSSK00032 94.6 5.4 2.6 uM 2.9 uM 30/36.5 nM NSSK00038 98.4 16.1 1.37uM 1.9 uM 30/36.5 nM NSSK00039 99.5 14.8 1.55 uM 870 nM 30/36.5 nMNSSK00041 99.3 13.3 1.1 uM 1.2 uM 30/36.5 nM NSSK00066 97.8 7.3 5.4 uM8.5 uM 30/36.5 nM NSSK00075 97.8 6.4 155 uM 653 uM 30/36.5 nM NSSK0004699.2 14.3 639 nM 518.9 nM 25.9/39.3 nM NSSK00047 99.8 4.5 355 nM 403.1nM 25.9/39.3 nM NSSK00073 98.5 3.6 119M 145.9M 25.9/39.3 nM NSSK0005699.4 15.7 752 nM 720 nM 25.9/39.3 nM NSSK00058 99.5 8.1 1.2 uM 1.5 uM25.9/39.3 nM NSSK00057 100.0 16.6 606 nM 795 nM 25.9/39.3 nM NSSK0006196.8 18 9.6 uM 10.2 uM 34/29.1 nM NSSK00049 97.7 14.5 2.8 uM 3.5 uM34/29.1 nM NSSK00051 99.5 10.1 1.1 uM 5.0 uM 34/29.1 nM NSSK00067 99.316.3  0.4M  0.2M 34/29.1 nM NSSK00042 98.6 4.3 134.7 nM 620.3 nM 34/29.1nM NSSK00059 95.3 5.8 1.57 uM 1.3 uM 34/29.1 nM NSSK00062 93.7 13 925.7nM 925.7 nM 43/43 nM NSSK00043 95.8 5.1 80.9 nM 80.7 nM 43/43 nMNSSK00044 96.3 13.8 149 nM 144.5 nM 43/43 nM NSSK00074 95.1 5.8 4.3 mM4.3 mM 43/43 nM NSSK00052 98.7 15.6 696.6 nM 696.6 nM 43/43 nM NSSK0004598.3 14.7 334 nM 334 nM 43/43 nM NSSK00064 94.4 6 4.6 uM 3.3 uM 117/120nM NSSK00072 96.1 12.7 1.1 uM 2.2 uM 117/120 nM NSSK00053 98.8 18.6529.4 nM 539 nM 117/120 nM NSSK00068 94.2 2.3 5.7 uM 7.4 uM 117/120 nMNSSK00060 98.4 4.2 8.5 uM 11.2 uM 117/120 nM NSSK00054 98.5 22.6 508.5nM 457 nM 117/120 nM NSSK00055 99.1 16.4 931 nM 1.4 uM 32/35 nMNSSK00048 98.4 14.2 382 nM 357 nM 32/35 nM NSSK00063 96.4 4.5 3 uM 1.9uM 32/35 nM NSSK00050 96.3 17.6 1.7 uM 2.1 uM 32/35 nM NSSK00065 94.2 103.3 uM 6.0 uM 32/35 nM NSSK00084 100.0 3 3.6 uM 4.7 uM 32/35 nMNSSK00087 94.3 2.2 1.2 mM 4.1 mM 292/287 nM NSSK00096 99.6 16.1 340 nM375 nM 292/287 nM (Salk 00096) NSSK00096 220 nM 63 nM (retested)NSSK00088 97.8 6.7 64 mM 34 mM 292/287 nM NSSK00089 100.0 16 383 nM 406nM 292/287 nM (Salk 00089) NSSK00089 366 nM 63 nM (retested) NSSK0009195.2 9.5 801 nM 628 nM 292/287 nM NSSK00097 99.7 3 866 nM 726 nM 292/287nM NSSK00095 97.6 4 1.4 uM 1.5 uM 51/56 nM NSSK00094 100.0 3.6 786 nM865 nM 51/56 nM NSSK00099 100.0 7.2 2.1 uM 2.1 uM 51/56 nM NSSK00098100.0 9.9 655 nM 670 nM 51/56 nM NSSK00100 100.0 10.1 1.4 uM 1.8 uM51/56 nM NSSK00092 95.7 5.7 3.4 uM 5.5 uM 51/56 nM NSSK00093 99.8 10.4459 nM 511 nM 81/88 nM NSSK00101 99.9 11.7 5.9 uM 15.1 uM 81/88 nMNSSK00077 96.5 12.5 177 nM 150 nM 81/88 nM (Deuterated Fexaramine)Deuterated 98 nM 63 nM Fexaramine (retested) NSSK00078 96.2 4.7 698.2 nM673 nM 81/88 nM NSSK00080 99.6 21.4 623 nM 610 nM 81/88 nM NSSK0008299.6 3.8 5.1 uM 6.1 uM 81/88 nM NSSK00081 98.3 6.1 22.8 uM 71 uM 77/100nM NSSK00079 97.3 19.6 513 nM 605 nM 77/100 nM NSSK00086 99.8 17.6 371nM 1.6 uM 97/320 nM NSSK00113 99.3 6.1 652 nM 2.7 uM 97/320 nM NSSK0007098.9 3.7 1.7 uM 1.7 uM 77/100 nM NSSK00102 98.8 2.2 252 nM 328 nM 77/100nM NSSK00107 97.5 11.6 450 nM 861 nM 77/100 nM NSSK00109 92.9 10.5 190nM 316 nM 77/100 nM NSSK00110 96.9 4 60.1 nM 174 nM 97/320 nM (Salk00110) NSSK00110 46 nM 63 nM (retested) NSSK00104 95.5 7.3 227 nM 547 nM97/320 nM NSSK00103 97.7 5.9 308 nM 1.0 uM 97/320 nM NSSK00114 92.7 12.2264 nM 614 nM 97/320 nM NSSK00108 93.7 19.5 not 497 nM 1/39 nM convergedNSSK00118 97.2 1.4 7 uM 55 uM 1/39 nM NSSK00119 98.3 1.9 798 nM 65 uM1/39 nM NSSK00115 98.4 5.4 1.4 mM 2.0 uM 1/39 nM NSSK00117 94.2 3.1 983nM 1.3 uM 1/39 nM NSSK00116 98.9 16.2 not 8.6 uM 1/39 nM converged

FIGS. 16-28 provide dose-response curves for exemplary compoundsindicating the relative activity and EC₅₀ values of the compounds.

Example 8 Synthesis of NSSK00110

8.1 General Procedure for Preparation of Compound 102

To the solution of compound 101 (13 g, 59 1 mmol) in triethylamine (TEA)(100 mL) was added methyl acrylate (54 mL, 591 mmol). Then the mixturewas degassed with N₂ for 5 minutes, and Pd(OAc)2 (2.6 g, 11.8 mmol) andtri(o-tolyl)phosphine (3.55 g, 11.8 mmol) were added and degassing wascontinued for another 5 minutes. The resulting solution was stirred atabout 90 to 100° C. under N₂ overnight. The solution was extracted withEtOAc (3×200 mL) and the combined organic extracts were washed withbrine (200 mL), dried over anhydrous Na₂SO₄, filtered, and concentratedin vacuo. The crude material was purified by Flash Chromatography(Petroleum ether/EtOAc=50:1) to give compound 102 (6.3 g, 48%) as awhite solid.

¹H NMR: (CDCl₃, 400 MHz) δ 8.19 (s, 1H), 7.94 (dt, J=5.6 Hz, 2.4 Hz,1H), 7.66 (d, J=16 Hz, 2H), 7.54 (dd, J=8.8 Hz, 2 Hz, 1H), 6.56 (d, J=16Hz, 1H), 3.84 (s, 3H).

8.2 General Procedure for Preparation of Compound 103

A mixture of compound 102 (9.8 g, 43.56 mmol) and SnCl₂.2H₂O (34 g,148.09 mmol) in anhydrous EtOH (150 mL) was heated at 80° C. for 2.5hours. Then the solvent was half removed under reduced pressure. Thesolution was poured into ice water and neutralized (pH=7) with saturatedNa₂CO₃ solution, then filtered and the filtrate was extracted with ethylacetate (3×200 mL). The combined organic extracts were washed with brine(200 mL), dried over anhydrous Na₂SO₄, filtered and concentrated invacuo to give compound 103 (7 g, 82.%) as a yellow solid. The productwas used directly in the next step without further purification.

¹H NMR: (CDCl₃, 400 MHz) δ7.52 (d, J=15.6 Hz, 1H), 6.62-6.50 (m, 2H),6.40-6.33 (m, 2H), 3.86 (brs, 2H), 3.80 (s, 3H).

8.3 General Procedure for Preparation of Compound 105

A 250 mL round-bottomed flask equipped with a Dean-Stark trap and refluxcondenser was charged with compound 103 (2.5 g, 13 mmol), compound 104(2.6 g, 13 mmol) and Tos-OH (300 mg, 1.7 mmol) in toluene (150 mL). Thesolution was refluxed at 130° C. for 48 hours until no more H₂O wascollected in the Dean-Stark trap. The volatiles were removed underreduced pressure to yield compound 105 (5 g, 100%) as a red solid. Theproduct was used directly in the next step without further purification.

TLC: Rf=0.5 (hexane/EtOAc:5/1)

8.4 General Procedure for Preparation of Compound 106

To a solution of compound 105 (5 g, 13 mmol) in THF (200 mL) was addedNaBD₄ (1.1 g, 26 mmol). The mixture was stirred at room temperature for16 hours. Then the mixture was quenched with saturated NH₄Cl solution,and the solution was extracted with ethyl acetate (3×100 mL) and thecombined organic layers were washed with brine (100 mL), dried overanhydrous Na₂SO₄, filtered, and concentrated in vacuo. The material waspurified by column chromatography on silica gel (petroleumether/EtOAc=3/1) to give compound 106 (2.9 g, 59%) as a yellow solid.

¹H NMR: (DMSO-d₆, 400 MHz) δ7.54 (dd, J=9.6, 2 Hz, 1H), 7.48 (d, J=16Hz, 1H), 7.42-7.37 (m, 1H), 7.36-7.30 (m, 1H), 6.77 (d, J=9.79 Hz, 1H),6.73 (d, J=1.51 Hz, 1H), 6.65 (d, J=5.77 Hz, 1H), 6.58-6.51 (m, 1H),6.42 (dt, J=11.67, 2.07 Hz, 1H), 4.35-4.28 (m, 1H), 3.71 (s, 3H).

8.5 General Procedure for Preparation of Compound 108

A solution of compound 107 (2.5 g, 14 mmol) in SOCl₂ (60 mL) was stirredat reflux for 3 hours under a nitrogen atmosphere. The mixture wasconcentrated in vacuo to give acid chloride as a yellow oil. To asolution of compound 106 (2.7 g, 7 mmol) in CH₂Cl₂ (60 mL) was added TEA(2.3 g, 21 mmol), followed by freshly-made acid chloride and DMAP (100mg, 0.8 mmol). The mixture was stirred at room temperature for 16 hours.Then the mixture was washed with water (50 mL), the aqueous layer wasextracted with CH₂Cl₂ (3×40 mL). The combined organic layers were washedwith brine (50 mL), dried over anhydrous Na₂SO₄, filtered, andconcentrated in vacuo. The material was purified by columnchromatography on silica gel (petroleum ether/EtOAc=30/1) to givecompound 108 (3.5 g, 97%) as a yellow solid.

¹H NMR: (DMSO-d₆, 400 MHz) δ7.68-7.52 (m, 3H), 7.44 (d, J=9.79 Hz, 1H),7.37-7.22 (m, 3 H), 6.76 (dd, J=16.19, 1.88 Hz, 1H), 5.04-4.70 (m, 1H),3.72 (s, 3H), 2.78 (brs., 0.5H), 2.34 (brs., 0.5H), 2.25-2.05 (m, 1H),1.89 (brs., 0.5H), 1.81-1.62 (m, 1H), 1.54 (brs., 0.5H), 1.48-1.15 (m,5H), 1.15-1.01 (m, 2H), 0.98-0.62 (m, 1H).

8.6 General Procedure for Preparation of NSSK00110

To a solution of compound 108 (3.5 g, 6 9 mmol) in dioxane/H₂O (100 mL)was added K₂CO₃ (2.8 g, 17.3 mmol) and Pd(dppf)Cl₂ (500 mg, 0.7 mmol),followed by compound 109 (1.5 g, 9 mmol). The mixture was stirred at 80°C. for 4 hours under a nitrogen atmosphere. The mixture was washed withwater (50 mL), the combined organic layers were washed with brine (10mL), dried over anhydrous Na₂SO₄, filtered, and concentrated in vacuo.The material was purified by prep-HPLC to give compound NSSK00110 (1.9g, 51%) as a yellow solid.

LCMS: MS (ESI) m/z 546 [M+H]⁺ (Purity: 100%)

¹H NMR: (DMSO-d₆, 400 MHz) δ7.68-7.57 (m, 2H), 7.56-7.48 (m, 3H),7.34-7.28 (m, 2H), 7.25 (d, J=9.13 Hz, 1H), 6.78-6.70 (m, 3H), 5.00-4.84(m, 1H), 3.71 (s, 3H), 2.92 (s, 6H), 2.80 (brs, 0.5H), 2.40 (brs, 0.5H),2.27-2.09 (m, 2H), 1.91 (brs, 0.5H), 1.83-1.65 (m, 1H), 1.58 (brs,0.5H), 1.49-1.19 (m, 3H), 1.07 (d, J=9.03 Hz, 2H), 0.94 (brs, 0.5H),0.71 (brs, 0.5H).

Example 9 Synthesis of NSSK00024

9.1 General Procedure for Preparation of Compound 111

A 250 mL round-bottomed flask equipped with a Dean-Stark trap and refluxcondenser was charged with compound 103 (5 g, 25 mmol), compound 110(4.7 g, 25 mmol) and Tos-OH (600 mg, 3.4 mmol) in toluene (200 mL). Thesolution was refluxed at 130° C. for 48 hours until no more H₂O wascollected in the Dean-Stark trap. The volatiles were removed underreduced pressure to yield compound 111 (9.7 g, 100%) as a red oil. Theproduct was used directly in the next step without further purification.

TLC: Rf=0.5 (hexane/EtOAc:5/1)

9.2 General Procedure for Preparation of Compound 112

To a solution of compound 111 (9.7 g, 25 mmol) in THF (200 mL) was addedNaBD₄ (2.2 g, 50 mmol), and the mixture was stirred at room temperaturefor 16 hours. Then the mixture was quenched with saturated NH₄Clsolution, and the solution was extracted with ethyl acetate (3×100 mL)and the combined organic layers were washed with brine (100 mL), driedover anhydrous Na₂SO₄, filtered, and concentrated in vacuo. The materialwas purified by column chromatography on silica gel(petroleum/EtOAc=3/1) to give compound 112 (5.6 g, 61%) as a yellowsolid.

¹H NMR: (DMSO-d₆, 400 MHz) δ7.52 (d, J=6.4 Hz, 2H), 7.46 (d, J=16 Hz,1H), 7.32 (d, J=8 Hz, 2H), 6.74-6.72 (m, 2H), 6.69 (s, 1H), 6.52 (d,J=16 Hz, 1H), 6.52 (d, J=12 Hz, 1H), 4.28-4.26 (m, 1H), 3.70 (s, 3H).

9.3 General Procedure for Preparation of Compound 113

A solution of compound 107 (3.2 g, 20 mmol) in SOCl₂ (60 mL) was stirredat reflux for 3 hours under a nitrogen atmosphere. The mixture wasconcentrated in vacuo to give acid chloride as a yellow oil. To asolution of compound 112 (3 g, 8 mmol) in CH₂Cl₂ (60 mL) was added TEA(2.5 g, 24 mmol), followed by fleshly acid chloride and DMAP (100 mg,0.8 mmol). The mixture was stirred at room temperature for 16 hours.Then the mixture was washed with water (50 mL), the aqueous layer wasextracted with CH₂Cl₂ (3×40 mL). The combined organic layers were washedwith brine (50 mL), dried over anhydrous Na₂SO₄, filtered, andconcentrated in vacuo. The material was purified by columnchromatography on silica gel (petroleum/EtOAc=30/1) to give compound 113(3.5 g, 97%) as a yellow solid.

¹H NMR: (DMSO-d₆, 400 MHz) δ7.61-7.57 (m, 2H), 7.50-7.40 (m, 3H),7.20-7.13 (m, 3H), 6.73 (d, J=16.4 Hz, 1H), 4.84-4.75 (m, 1H), 3.72 (s,3H), 2.78 (brs, 0.5H), 2.34 (brs, 0.5H), 2.25-2.12 (m, 2H), 2.0-1.2 (m,5H), 1.15-1.01 (m, 2H), 0.98-0.60 (m, 1H).

9.4 General Procedure for Preparation of NSSK00024

To a solution of compound 113 (3.5 g, 8 2 mmol) in dioxane/H₂O (100 mL)was added K₂CO₃ (3.4 g, 20.5 mmol) and Pd(dppf)Cl₂ (600 mg, 0.8 mmol),followed by compound 109 (1.7 g, 10.6 mmol). The mixture was stirred at80° C. for 4 hours under a nitrogen atmosphere. The mixture was washedwith water (50 mL), the combined organic layers were washed with brine(10 mL), dried over anhydrous Na₂SO₄, filtered, and concentrated invacuo. The material was purified by prep-HPLC to give compound NSSK00024(2.3 g, 53%) as a yellow solid.

LCMS: MS (ESI) m/z 528 [M]H⁺ (Purity: 99%)

¹H NMR: (DMSO-d₆, 400 MHz) δ7.61 (d, J=16.06 Hz, 2H), 7.48 (t, J=8.28Hz, 5H), 7.19 (dd, J=8.78, 2.51 Hz, 3H), 6.79-6.69 (m, 3H), 5.00-4.77(m, 1H), 3.71 (s, 3H), 2.91 (s, 6H), 2.86(brs, 0.5H), 2.37 (brs., 0.5H),2.30-2.09 (m, 2H), 1.92 (brs, 0.5H), 1.67-1.84 (m, 1H), 1.60 (brs,0.5H), 1.52-1.19 (m, 3H), 1.08 (d, J=9.29 Hz, 2H), 0.95 (brs, 0.5H),0.72 (brs, 0.5H).

Example 10 Synthesis of NSSK000027

10.1 General Procedure for Preparation of Compound 116

To a mixture of compound 115 (82.4 g, 452 mmol) and K₂CO₃ (96 g, 696mmol) in H₂O (160 mL) was added compound 114 (52.4 g, 348 mmol). Thereaction was stirred at room temperature for 1 hour. Then the solutionwas filtered and the filter cake was washed with 1 N HCl and water, andthe solid was concentrated to give compound 116 (85 g, crude) as a whitesolid. The product was used directly in the next step without furtherpurification.

¹H NMR: H20619-001-1Q1 (DMSO-d₆, 400 MHz) δ8.56 (s, 1H), 8.25-8.19 (m,2H), 7.80 (d, J=16 Hz, 1H), 7.71 (t, J=8 Hz, 1H), 6.86 (d, J=16 Hz, 1H),3.75 (s, 3H).

10.2 General Procedure for Preparation of Compound 117

A mixture of compound 116 (50 g, 241.5 mmol) and SnCl₂.2H₂O (186.3 g,820.7 mmol) in anhydrous EtOH (600 mL) was heated at 80° C. for 2.5hours. Then the solvent was half removed under reduced pressure. Thenthe solution was poured into ice water and neutralized (pH=7) withsaturated Na₂CO₃ solution, and the solution was extracted with ethylacetate (3×500 mL) and the combined organic layers were washed withbrine (500 mL), dried over anhydrous Na₂SO₄, filtered, and concentratedin vacuo to give compound 117 (32.3 g, 75.7%) as a yellow solid. Theproduct was used directly in the next step without further purification.

¹H NMR: (CDCl₃, 400 MHz) δ7.60 (d, J=15.6 Hz, 1H), 7.17 (t, J=7.6 Hz,1H), 6.93 (d, J=7.6 Hz, 1H), 6.82 (s, 1H), 6.71 (dd, J=8 Hz, 1.5 Hz,1H), 6.38 (d, J=15.6 Hz, 1H), 3.80 (s, 3H), 3.76 (brs, 2H).

10.3 General Procedure for Preparation of Compound 118

A 250 mL round-bottomed flask equipped with a Dean-Stark trap and refluxcondenser was charged with compound 117 (5 g, 28.25 mmol), compound 104(5.75 g, 28.25 mmol) and Tos-OH (600 mg, 0.58 mmol) in toluene (100 mL).The solution was refluxed at 110° C. for 24 hours until no more H₂O wascollected in the Dean-Stark trap. The volatiles were removed underreduced pressure to yield compound 118 (10.2 g, 100%) as red oil, whichwas sent to next step directly without further purification.

TLC: Rf=0.8 (Petroleum ether/EtOAc=3/1)

10.4 General Procedure for Preparation of Compound 119

To a solution of compound 118 (10.2 g, 28.25 mmol) in THF (135 mL) wasadded NaBD₄ (2.37 g, 56.5 mmol), and the mixture was stirred at roomtemperature for 16 hours. The mixture was quenched with saturated NH₄Clsolution, and the solution was extracted with ethyl acetate (3×120 mL)and the combined organic layers were washed with brine (120 mL), driedover anhydrous Na₂SO₄, filtered, and concentrated in vacuo. The materialwas purified by Flash Chromatography (Petroleum ether: EtOAc=30:1) togive compound 119 (4.6 g, 45.1%) as a yellow solid. ¹H NMR: (DMSO-d₆,400 MHz) δ7.54-7.48(m, 2H), 7.37-7.33 (m, 2H), 7.11 (t, J=7.6 Hz, 1H),6.89-6.86 (m, 2H), 6.65 (dd, J=8 Hz, 1.6 Hz, 1H), 6.47 (d, J=16 Hz, 1H),6.35 (d, J=6.4 Hz, 1H).

10.5 General Procedure for Preparation of Compound 121

To a solution of compound 119 (5.6 g, 15.33 mmol) in CH₂Cl₂ (130 mL) wasadded compound 120 (4.5 g, 30.67 mmol), TEA (4.65 g, 45.99 mmol) andDMAP (280 mg, 2.30 mmol). The mixture was stirred at room temperaturefor 1 hour. Then the mixture was washed with water (3×100 mL), thecombined organic layers were washed with brine (100 mL), dried overanhydrous Na₂SO₄, filtered, and concentrated in vacuo, then purified bypre-HPLC to give compound 121 (5.7 g, 70%) as a white solid.

¹H NMR: (DMSO-d₆, 400 MHz) δ7.71-7.61(m, 3H), 7.44-7.40 (m, 2H), 7.35(dd, J=8 Hz, 2 Hz, 1H), 7.26 (t, J=8 Hz, 1H), 7.19 (d, J=8 Hz, 1H), 4.84(br. s., 1H) 3.72 (s, 3H), 2.20-2.08 (br. s., 1H), 1.69-1.55 (m, 4H),1.50-1.47 (m, 1H) 1.42-1.30 (m, 2H), 1.15-1.03 (m, 1H), 0.94-0.80 (m,2H).

10.6 General Procedure for Preparation of NSSK00027

To a solution of compound 121 (2.3 g, 4.84 mmol) in dioxane/H20 (70 mL)was added K₂CO₃ (2 g, 14.52 mmol) and Pd(dppf)Cl₂ (357 mg, 0.48 mmol),followed by compound 109 (1.04 g, 6.29 mmol). The mixture was stirred at80° C. for 4 hours under a nitrogen atmosphere. The reaction mixture wasfiltered and the filtrate was extracted with ethyl acetate (3×70 mL) andthe combined organic layers were washed with brine (70 mL), dried overanhydrous Na₂SO₄, filtered, and concentrated in vacuo. The material waspurified by Flash Chromatography (Petroleum ether: EtOAc=8:1) to givecrude product. The crude material was purified by prep-HPLC to giveNSSK00027 (1.8 g, 72%) as a green solid.

LCMS: MS (ESI) m/z=516 [M]H⁺ (Purity: 100%)

¹H NMR: (DMSO-d₆, 400 MHz) δ7.72-7.61 (m, 3H), 7.52 (d, J=8.8 Hz, 2H),7.45-7.36 (m, 2H), 7.33-7.26 (m, 2H), 7.20 (d, J=7.9 Hz, 1H) 6.76 (d,J=8.8 Hz, 2H), 6.67 (d, J=16 Hz, 1H), 4.88 (brs, 1H), 3.71 (s, 3H), 2.93(s, 6H), 2.22-2.10 (m, 1H) 1.71-1.57 (m, 4H), 1.54-1.46 (m, 1H)1.45-1.33 (m, 2H), 1.16-1.06 (m, 1H), 0.96-0.80 (m, 2H).

Example 11 Synthesis of NSSK00089

To a solution of compound 121 (2.3 g, 4.84 mmol) in dioxane/H₂O (70 mL)was added K₂CO₃ (2 g, 14.52 mmol) and Pd(dppf)Cl₂ (357 mg, 0.48 mmol),followed by compound 122 (1.1 g, 6.29 mmol). The mixture was stirred at80° C. for 4 hours under a nitrogen atmosphere. The reaction mixture wasfiltered and the filtrate was extracted with ethyl acetate (3×70 mL) andthe combined organic layers were washed with brine (70 mL), dried overanhydrous Na₂SO₄, filtered, and concentrated in vacuo. The material waspurified by Flash Chromatography (Petroleum ether: EtOAc=8:1) to givecrude product. The crude material was purified by prep-HPLC to giveNSSK00089 (1.9 g, 74%) as a white solid.

LCMS: MS (ESI) m/z=527 [M+H]⁺ (Purity: 99%)

¹H NMR: (DMSO-d₆, 400 MHz) δ8.06 (d, J=17.2 Hz, 2H), 7.76-7.59 (m, 5H),7.53-7.35 (m, 4H), 7.24 (d, J=8 Hz, 1H), 6.67 (d, J=16 Hz, 1H), 4.93(br. s., 1H) 4.06 (s, 3H), 3.70 (s, 3H), 2.23-2.12 (m, 1H), 1.74-1.56(m, 4H), 1.54-1.34 (m, 3H), 1.18-1.03 (m, 1H), 0.98-0.79 (m, 2H).

Example 12 Synthesis of NSSK00096

12.1 General Procedure for Preparation of Compound 123

To a solution of compound 112 (2.5 g, 6.8 mmol) in CH₂Cl₂ (60 mL) wasadded TEA (2 g, 20.4 mmol), followed by compound 120 (2 g, 13.7 mmol),and DMAP (248 mg, 2 mmol). The mixture was stirred at room temperaturefor 3 hours. Then the mixture was washed with water (50 mL), the aqueouslayer was extracted with CH₂Cl₂ (3×40 mL). The combined organic layerswere washed with brine (50 mL), dried over anhydrous Na₂SO₄, filtered,and concentrated in vacuo. The material was purified by columnchromatography on silica gel (petroleum/EtOAc=30/1) to give compound 123(2.4 g, 75%) as a yellow solid.

¹H NMR: (DMSO-d₆, 400 MHz) δ7.66-7.58(m, 2H), 7.47-7.45 (m, 3H),7.18-7.12 (m, 3H), 6.74 (d, J=15.6 Hz, 1H), 4.81 (s., 1H), 3.72 (s, 3H),2.20 (brs, 1H), 1.71-1.59 (m, 4H), 1.52-1.49 (m, 1H), 1.42-1.15 (m, 2H),1.12-1.09 (m, 1H), 0.94-0.91 (m, 2H).

12.2 General Procedure for Preparation of NSSK00096

To a solution of compound 123 (2.4 g, 5 mmol) in dioxane/H₂O (50 mL) wasadded K₂CO₃ (2 g, 12.5 mmol) and Pd(dppf)Cl₂ (366 mg, 0.5 mmol),followed by compound 122 (1.4 g, 6.5 mmol). The mixture was stirred at80° C. for 4 hours under a nitrogen atmosphere. The mixture was washedwith water (50 mL), the combined organic layers were washed with brine(10 mL), dried over anhydrous Na₂SO₄, filtered, and concentrated invacuo. The material was purified by prep-HPLC to give compound NSSK00096(1.1 g, 42%) as a yellow solid.

LCMS: MS (ESI) m/z=527 [M+H]⁺ (Purity: 100%)

¹H NMR: (DMSO-d₆, 400 MHz) δ8.07 (s, 1H), 7.97 (d, J=0.98 Hz, 1H),7.72-7.58 (m, 6H), 7.50 (s, 1H), 7.26 (d, J=8.07 Hz, 2H), 7.20 (dd,J=9.41, 1.83 Hz, 1H), 6.73 (d, J=15.90 Hz, 1H), 4.90 (s., 1H), 4.05 (s,3H), 3.71 (s, 3H), 2.24 (brs, 1H), 1.73 (d, J=11.74 Hz, 2H), 1.63 (d,J=12.72 Hz, 2H), 1.52 (d, J=11.74 Hz, 1H), 1.48-1.33 (m, 2H), 1.12 (q,J=12.72 Hz, 1H), 0.96 (brs, 2H).

Example 13 Synthesis of NSSK00077

13.1 General Procedure for Preparation of Compound 124

A 250 mL round-bottomed flask equipped with a Dean-Stark trap and refluxcondenser was charged with compound 117 (3 g, 16.9 mmol), compound 110(3.09 g, 16.9 mmol) and Tos-OH (348 mg, 2.02 mmol) in toluene (100 mL).The solution was refluxed at 110° C. for 24 hours until no more H₂O wascollected in the Dean-Stark trap. The volatiles were removed underreduced pressure to yield compound 124 (5.8 g, 100%) as red oil, whichwas sent to next step directly without further purification.

TLC: Rf=0.8 (Petroleum ether/EtOAc=3/1)

13.2 General Procedure for Preparation of Compound 125

To a solution of compound 124 (5.8 g, 16.9 mmol) in THF (100 mL) wasadded NaBD₄ (1.42 g, 33.8 mmol). The mixture was stirred at roomtemperature for 16 hours. Then the mixture was quenched with saturatedNH₄Cl solution, and the solution was extracted with ethyl acetate (3×100mL) and the combined organic layers were washed with brine (100 mL),dried over anhydrous Na₂SO₄, filtered, and concentrated in vacuo. Thematerial was purified by Flash Chromatography (Petroleum ether:EtOAc=30:1) to give compound 125 (3.6 g, 62.1%) as a yellow solid.

¹H NMR: (DMSO-d₆, 400 MHz) δ7.55-7.45 (m, 3H), 7.32 (d, J=8.4 Hz, 2H),7.09 (t, J=7.6 Hz, 1H), 6.88-6.82 (m, 2H), 6.63 (d, J=8 Hz, 1H),6.47-6.40 (m, 2H), 4.26 (d, J=6.36 Hz, 1H), 3.70 (s, 3H).

13.3 General Procedure for Preparation of Compound 126

To a solution of compound 125 (3.6 g, 10 4 mmol) in CH₂Cl₂ (100 mL) wasadded compound 120 (3.16 g, 21.49 mmol), TEA (3.27 g, 32.34 mmol) andDMAP (197 mg, 1.62 mmol). The mixture was stirred at room temperaturefor 1 hour. Then the mixture was washed with water (3×100 mL), thecombined organic layers were washed with brine (100 mL), dried overanhydrous Na₂SO₄, filtered, and concentrated in vacuo, then purified bypre-HPLC to give compound 126 (4.3 g, 90%) as a white solid.

¹H NMR: (DMSO-d₆, 400 MHz) δ7.73-7.60 (m, 3H), 7.50-7.38 (m, 3H),7.17-7.08 (m, 3H), 6.68 (d, J=16.4 Hz, 1H), 4.80 (br. s., 1H), 3.72 (s,3H), 2.20-2.08 (m, 1H), 1.72-1.56 (m, 4H), 1.54-1.46 (m, 1H), 1.45-1.31(m, 2H), 1.17-1.04 (m, 1H) 0.95-0.82 (m, 2H).

13.4 General Procedure for Preparation of NSSK00077

To a solution of compound 126 (2.5 g, 5.47 mmol) in dioxane/H₂O (70 mL)was added K₂CO₃ (2.27 g, 16.41 mmol) and Pd(dppf)Cl₂ (412 mg, 0.55mmol), followed by compound 109 (1.18 g, 7.12 mmol). The mixture wasstirred at 80° C. for 4 hours under a nitrogen atmosphere. The mixturewas then filtered and the filtrate was extracted with ethyl acetate(3×70 mL) and the combined organic layers were washed with brine (70mL), dried over anhydrous Na₂SO₄, filtered, and concentrated in vacuo.The material was purified by Flash Chromatography (Petroleum ether:EtOAc=8:1) to give crude product. The crude material was purified byprep-HPLC to give NSSK00077 (1.9 g, 71%) as a yellow solid.

LCMS: MS (ESI) m/z=498 [M+H]⁺ (Purity: 99%)

¹H NMR: H20619-033-1H5 (DMSO-d₆, 400 MHz) δ7.72-7.59 (m, 3H), 7.53-7.44(m, 4H), 7.41 (t, J=8 Hz, 1H), 7.16 (d, J=7.6 Hz, 2H), 6.77 (d, J=8.8Hz, 2H), 6.66 (d, J=15.6 Hz, 1H), 4.84 (br. s., 1H), 3.71 (s, 3H), 2.91(s, 6H), 2.24-2.12 (m., 1H), 1.73-1.57 (m, 4H), 1.55-1.34 (m, 3H),1.17-1.04 (m, 1H), 0.97-0.79 (m, 2H).

Example 14 Effect of Fexaramine and Selectively-deuterated FexaramineAnalogs in vivo

An in vivo study of fexaramine and selectively-deuterated fexaraminecompounds (FIG. 29A) was performed. Briefly, ob/ob mice were treateddaily with vehicle, Fex, or deuterated analogs (50 mg kg⁻¹) by oral (PO)gavage for 2 weeks. Subsequently, body weight, body temperature, fastingblood glucose and insulin levels, insulin secretion, and GLP1 sectionwere measured, and glucose tolerance tests (GTTs) performed.

As shown in FIGS. 29B-29C, treatment with fexaramine or a fexaramineanalog treatment (2 weeks at 50 mg/kg) of ob/ob mice do not affect bodyweight, but did increase core body temperature. Fex-D treated was moreeffective at increasing core body temperature than Fex. DeuteratedSALK110 (NSSK00110) showed superior activation compared to fexaramine

As shown in FIG. 29D, while two weeks treatment with fexaramine does notsignificantly affect fasting blood glucose levels, FEX-D and SALK110(NSSK00110) treatment significantly reduced fasting blood glucoselevels.

As shown in FIG. 29E, GTTs performed after two weeks of treatment withthe indicated analogs demonstrated improved glucose tolerance with thedeuterated analogs, but not with fexaramine, demonstrating superioractivity with the deuterated analogs.

As shown in FIG. 29F, the deuterated fexaramine analogs were more activethan fexaramine at lowering fasting insulin levels.

As shown in FIG. 29G, treatment with the deuterated fexaramine analogsincreased insulin secretion in ob/ob mice in response to a glucosechallenge, as measured during the GTT.

As shown in FIG. 29H, GLP1secretion in ob/ob mice in response to aglucose challenge in mice treated with the deuterated fexaramineanalogs, as measured during the GTT, increased GLP1 secretion whichleads to increased insulin secretion.

Example 15 Orally Delivered Fexaramine Analogs are Intestinal-specificFXR Agonists

Mice were treated daily with vehicle, FEX-D, or SALK110 (NSSK00110, 50mg kg⁻¹) (see FIG. 29A) by oral (PO) gavage for 14 days, with tissuescollected 1 hour after the final treatment. Gene expression changes inthe liver were measured by QPCR.

As shown in FIG. 30, FEX-D and SALK110 (NSSK00110) fail to alter theexpression of the canonical FXR target gene SHP and BSEP in the liver,indicating that both analogs are intestinal-specific (they do not enterthe circulation when delivered orally). However, activation of FXR inthe intestine induces the paracrine factor Fgf15, as demonstrated byreduced levels of Fgf15 target genes in the liver.

Example 16 Co-Upregulated Genes in Islets by Chronic Treatment ofFexaramine Analogs

Pancreatic islets were isolated from ob/ob mice after daily treatment byoral gavage with fexaramine (100 mg/kg for 5 weeks), FEX-D (50 mg/kg for2 weeks) or SALK110 (NSSK00110, 50 mg/kg for 2 weeks). Changes in geneexpression were determined by RNA-Seq.

As shown in FIG. 31 and Table 6, fexaramine and the deuterated analogsinduce a common set of genes in islets that are involved inintracellular signaling, insulin secretion, and regulation ofexocytosis. These gene changes are consistent with the increased insulinsecretion seen in FIG. 29G.

TABLE 6 The functional annotation of common gene expression changesinduced by fexaramine analogs, as determined by gene ontology. GeneFunctions P value Intraceullular signaling casade 3.2E−02 Insulinsecretion 5.2E−02 Regulation of exocytosis 6.2E−02 Peptide hormonesecretion 6.5E−02

Example 17 Intestinal Permeability Assay of Fexaramine Analogs

Standard Caco-2 cell permeability assays were performed on fexaramineand analogs as an in vitro evaluation of their intestinal permeability.The assays were performed as follows. Briefly, standard Caco-2 CultureMedia DMEM FCS 10% L-Glutamine 1% PenStrep 1% (sterile-filtered) wasplaced in CacoReady 24 well transwell plate, obtained from ADMEcell(Alameda, Calif.). The plates were incubated in a 37° C., 5% CO₂incubator for 4 hours. About 5 ml of 1000-fold diluted compound solutionin transport buffer was prepared. The basal assay plate was prepared byadding 750 μl of transport buffer to A-B wells, and 780 μl of dilutedcompound solution to B-A wells. The basal assay plate was placed in theincubator. The CacoReady plate was put into a hood, apical section ofplate lifted out and lowered onto empty basal plate. 200 μl of theCoco-2 media was removed from the apical wells and replaced with 200 μlof fresh transport media. This was repeated twice for a total of 3washes. 200 μl of the media was removed from the apical wells andreplaced with 200 μl of diluted compound (for A-B wells) or 200 μl offresh transport buffer (for B-A wells). The basal plate was removed fromthe incubator and the apical section of plate transferred to the basalplate. Three replicate, 10 μl samples were collected from the apical andbasal compartments for T0. The assay plate was covered and returned tothe incubator. At T2 hrs, 3 replicate, 10 μl samples were collected fromall apical compartments and B-A basal compartments; 3 replicate, 50 μlsamples were collected from A-B basal compartments. 50 μl of all T0 andT2hrs samples were mixed and transferred for bioanalysis.

Calculations: Analyte levels (peak area ratios) are measured on apical(A) and basolateral (B) sides at T0 and T2hrs. A-to-B and B-to-A fluxesare calculated (mean of n=3 measurements). Apparent permeability (Papp,cm/sec) is calculated as dQ (flux)/(dt×Area×Concentration). The effluxratio is (B-to-A)/(A-to-B) ratio [i.e., Papp(B-A)/Papp(B-A)]. A ratio >2is evidence of efflux. PGP efflux can be confirmed by testing +/− pgpinhibitor (dosing solutions prepared with and without verapamil at afinal assay concentration of 25 μM).

As shown in FIG. 32, all analogs are poorly transported, consistent withthese molecules being intestinally restricted.

Example 18 Intestinal Activation of the Nuclear Receptor FXR viaFexaramine Improves Tumor Burden in an APC^(min) Colon Cancer Mice ModelBackground

The lining of the mammalian intestine is comprised of a rapidlyproliferating epithelial monolayer that undergoes continuous renewal. Itprovides two vital functions: absorbing nutrients and water, and servingas a physical barrier that separates the immune system from luminalbacteria, antigens, and toxins. Tissue homeostasis of the adultintestinal epithelium depends on cellular plasticity to enableself-renewal, proliferation, and apoptosis, and to ensure effectivewound healing without promoting malignant outgrowth. These processes areregulated in part through Wnt-β-catenin signaling, which promotesproliferation of the epithelial stem cell compartment at the base of theintestinal crypt and is required for intestinal regeneration. Wntligands are secreted glycoproteins that activate the Frizzled family ofG protein (heterotrimeric guanine nucleotide-binding protein)-coupledreceptors and the co-receptors Lrp5 and Lrp6. Activation of Wnt receptorcomplexes leads to inhibition of a protein complex, which includes thetumor suppressor protein APC (adenomatous polyposis coli), whichpromotes ubiquitin-mediated degradation of the transcriptionalcoactivator β-catenin. In greater than 80% of sporadic and familialcolorectal cancers (CRCs), mutations causing premature stop codons inAPC induce constitutive accumulation and activation of β-catenin in thenucleus and drive tumor formation. Therefore, therapies designed tointerfere with the Wnt-β-catenin pathway in CRC are of clinical use.

Methods

The farnesoid X receptor (FXR) is a bile acid activated nuclear receptorwidely expressed through the body. The ability of theintestinal-specific FXR ligand fexaramine to inhibit the progression ofcolon cancer in mice with a heterozygote mutation for the Apc gene(ApcMin/+ mice on a C57B6 background obtained from Jackson Laboratories)was determined Mice were maintained under temperature, air andlight-controlled conditions and received food and water ad libitum; theydid not receive any surgical or hormonal manipulation.

5 week old ApcMin/+ mice (13-15 males per group receiving a standarddiet, a model for colorectal and intestinal cancer) were treated withvehicle (corn oil) or fexaramine (FEX, 100 mg/kg in corn oil) by oralgavage, three times a week for 23 weeks. Mice body weight andfood-intake were measured daily. At the end of the treatment regimen,surviving animals were sacrificed by cervical dislocation and the entireintestinal tract was immediately removed and washed with coldphosphate-buffered saline.

Tumor analysis, histological scores and colonic length At the completionof the treatment, the colons were removed, flushed with PBS, fixed as“Swiss-rolls” in 4% paraformaldehyde at 4° C. overnight, andparaffin-embedded. Sections (5 μm) were cut stepwise (200 μm) throughthe complete block and stained with H&E. Tumor counts were performedblinded by a trained pathologist. To determine proliferation rates, micewere injected i.p. with 100 mg/kg of BrdU 2.5 hours prior to sacrificeand paraffin sections stained using a BrdU-in situ detection kit. Theextent of apoptosis was determined by TUNEL assay using the ApoAlert DNAfragmentation assay kit. BrdU- and TUNEL-positive cells were counted byan investigator blinded to the genotype. For immunohistochemistry,antigen retrieval was performed (Vector Lab H3300) and then RTUVectastain Universal Elite ABC Kit and NovaRED or VIP substrate (allfrom Vector Lab) were used, following the manufacturer's instructions.

Statistical analysis Statistical analysis were performed incollaboration with the Cancer Prevention-Biostatistics department at theUniversity of California at San Diego Cancer Center. This is part of anongoing collaboration and has previously been applied to the APC^(Min)model of colorectal cancer as well as to several other cancer modelsincluding prostate and breast cancer. The volume of polyps wascalculated considering them as hemispheres (½×¾ πr³). All otherevaluations were performed in the distal tract of the small intestinebecause ApcMin/+ mice develop the majority of tumors in the smallintestine.

Results

As shown in FIG. 33A, fexaramine-treated mice are resistant to coloncancer-induced cachexia (weight loss). As shown in FIG. 33B,vehicle-treated mice lose 10% body weight, while Fex-treated micemaintain their weight. As shown in FIG. 33C, a dramatic improvement insurvival of Fex-treated mice compared to vehicle-treated (vehicle 20%survival at 25 weeks vs. 80% for Fex treated mice) was observed.

As shown in FIG. 34A, a dramatic reduction in tumor burden ofFex-treated versus vehicle-treated mice was observed after 23 weeks (20%reduction, p 0.04). As shown in FIG. 34B, no change in tumor burden ofvehicle- versus FEX-treated mice was observed in the duodenum after 23weeks (p 0.73). As shown in FIG. 34C, a dramatic reduction in tumorburden in Fex-treated versus vehicle-treated treated mice was observedin the jejunum after 23 weeks (11.75% reduction, p 0.08). As shown inFIG. 34D, a dramatic reduction in tumor burden in Fex-treated versusvehicle-treated treated mice was observed in the ileum after 23 weeks(25% reduction, p 0.08)

As shown in FIGS. 35A-35D, a dramatic reduction in tumor size inFex-treated versus vehicle-treated treated mice was observed in theduodenum (FIG. 35A), the jejunum (FIG. 35B),the ileum (FIG. 35C), andthroughout the intestine (FIG. 35D) after 23 weeks.

As shown in FIG. 36, serum total cholesterol and triglyceride levels inAPC^(min) mice after 23 weeks is reduced in fexaramine-treated, but notvehicle-treated mice. Very high triglycerides are dangerous. Levelsabove 500 mg/dL can cause fatty deposits in the skin and internalorgans, which can damage the liver and pancreas.

As shown in FIGS. 37-40, respectively, duodenum, jejunum, ileum andcolon paraformaldehyde fixed intestinal sections of APC^(min) micetreated with fexaramine have reduced tumor size as compared tovehicle-treated mice.

In summary, fexaramine-treated APC^(min) mice are resistant to cachexia,have an improved survival rate, have delayed tumor progression, and havereduced tumorigenesis (e.g., fewer tumor counts in ileum). In addition,the severe tumor burdens positively correlates with serum turbidity invehicle group. Thus, use of fexaramine, or any of the fexaraminederivatives thereof provided herein, can be used to treat cancer, suchas colon cancer for example by reducing one or more of cachexia, numberof tumors, metastasis, and size of tumor(s), such as a reduction of atleast 5%, at least 10%, at least 15%, at least 20%, or at least 50%,and/or increasing survival rate, such as an increase of at least 5%, atleast 10%, at least 15%, at least 20%, or at least 50%.

Example 19 Administration of Fexaramine and Guggulsterone

Glucagon-like peptide 1 (GLP-1) is a gut-derived peptide secreted byintestinal L cells after a meal, where it functions to potentiateglucose-stimulated insulin secretion, enhance β-cell growth andsurvival, and inhibit gastric emptying and food intake. The demonstratedglucose-lowering effects of GLP-1 have lead to the approval of GLP-1receptor agonists for the treatment of Type 2 diabetes. However, GLP-1secretion is reduced in patients with type 2 diabetes, leading tointerest in GLP-1 secretagogues as alternative therapies.

To examine the effects of FXR in the secretion of GLP-1, the metabolicchanges induced in human L cells was measured by treatment with the FXRagonist, fexaramine and an FXR antagonist guggulsterone. Treatment of Lcells with fexaramine (1 μM for 24 hours) lead to an increase in theoxygen consumption rate (OCR), consistent with increased mitochondrialactivity and consequently, an increased energetic state (FIG. 41). Thereverse effect was seen after treatment with the FXR antagonist,guggulsterone, with lower OCR after drug treatment. The ability offexaramine to increase the energetic state of the L cells indicates thatit can function as a GLP-1 secretagogue.

Example 20 Administration of Fexaramine Protects from Alcoholic LiverDisease

Patients with alcoholic hepatitis have a high mortality rate. Inaddition, patients with alcoholic liver disease show an overgrowth ofintestinal bacteria.

The Tsukamoto-French mouse model, which involves continuous intragastricfeeding of isocaloric diet or ethanol for 3 weeks, was used. C57BL/6Jmice were co-administered vehicle or fexaramine by oral gavage (100mg/kg/day). Mice were sacrificed after treatment and conjugated andunconjugated bile acid levels measured in serum and liver by liquidchromatography/mass spectrometry.

As shown in FIGS. 42A-42C, increased choloylglycine hydrolase activityis seen in the liver of alcoholic patients. Choloylglycine hydrolase isresponsible for the deconjugation of bile acids. Ethanol treatment ofmice leads to an increase in the level of deconjugated bile acids inserum, and an increase in the level of conjugated bile acids in theliver, as seen in human patients. In addition, there is a markedincrease in the total bile acid levels. Consistent with this increase intotal bile acid levels, ethanol treatment increased the expression ofCyp7a1, the enzyme that catalyzes the rate limiting step in theconversion of cholesterol to bile acids in the liver (FIGS. 43A and43B).

As shown in FIG. 44A, co-administration of fexaramine to mice during theadministration of ethanol protects them from alcoholic liver disease, bydecreasing hepatic steatosis, as shown histologically (FIG. 44A) andquantified in FIG. 44C Improved liver function is also indicated bydecreased serum levels of alanine aminotransferase (ALT, FIG. 44B).

In view of the many possible embodiments to which the principles of thedisclosure may be applied, it should be recognized that the illustratedembodiments are only examples of the disclosure and should not be takenas limiting the scope of the invention. Rather, the scope of theinvention is defined by the following claims. We therefore claim as ourinvention all that comes within the scope and spirit of these claims.

We claim:
 1. A compound, having a formula

or a pharmaceutically acceptable salt, hydrate, solvate or prodrugthereof, wherein R¹ is selected from aryl, heteroaryl, heterocyclic,alkenyl, cycloalkyl, cycloalkenyl or polycyclic; R² is selected fromalkyl, alkenyl, or cycloalkyl; Y is selected from N, N—O or C—R^(3d);R^(3a), R^(3b), R^(3c) and R^(3d) are each independently selected fromhydrogen, deuterium, halide, alkyl, alkenyl, alkoxy, alkylthio, amino,sulfonyl, aminosulfonyl, aminocarbonyl, acyl, hydroxyl or nitro; R^(4a)and R^(4b) are each independently selected from hydrogen, deuterium,halide or alkyl; L¹ and L² are independently selected from hydrogen,deuterium, alkyl, cycloalkyl, or together form a pi-bond; and R^(5a),R^(5b), R^(5c), R^(5d) and R^(5e) are each independently selected fromhydrogen, deuterium, halide, alkyl, alkenyl, alkoxy, alkylthio, amino,sulfonyl, aminosulfonyl, aminocarbonyl, acyl, aryl, heteroaryl,cycloalkyl, heterocyclyl, hydroxyl or nitro; or any two adjacent groupsselected together form an aryl, heteroaryl, cycloalkyl or heterocyclicring; and wherein none of R^(5a), R^(5b), R^(5c), R^(5d) or R^(5e) are—R^(x)-L^(x)-R^(x2), where R^(x) is selected from O, NR^(x3), sulfonylor S; R^(x3) is selected from H, alkyl, alkenyl, alkynyl, cycloalkyl,cycloalkenyl, or aryl; L^(x) is selected from a bond, alkylene,alkenylene, alkynylene, cycloalkyl, cycloalkenyl, heterocyclic, aryl,heteroaryl or CR^(x4)R^(x5); R^(x4) and R^(x5) are each independentlyselected from H, D, halogen, alkyl, alkenyl, alkynyl, cycloalkyl,cycloalkenyl, —C(O)OR^(x6), or C(O)NR^(x6)R^(x7); R^(x6) and R^(x7) areeach independently selected from H, alkyl, alkenyl, alkynyl, cycloalkylor cycloalkenyl; R^(x2) is selected from —C(O)L^(x2)R^(x8) or a carboxylbioisostere; L^(x2) is a bond or NR^(x3); R^(x8) is H, alkyl, alkenyl,alkynyl, cycloalkyl, cycloalkenyl, —OR^(x9), N(R^(x9))₂, —C(O)R^(x9),—S(O)₂R^(x9), —C(O)OR^(x9), —S(O)₂N(R^(x9))₂ or —C(O)N(R^(x9))₂; andeach R^(x9) is independently selected from H, alkyl, alkenyl, alkynyl,cycloalkyl or cycloalkenyl; and if L¹ and L² are both hydrogen ortogether form a pi-bond then Y is N or C-halogen; or R¹ is polycyclic;or R^(4a) is D; or R^(5a) is F, Cl or I; or R^(5d) and R^(5e) togetherform an aryl, heteroaryl, cycloalkyl or heterocyclic ring; or R^(5b) andR^(5e) together form an aryl, cycloalkyl, nitrogen-containingheterocyclic or nitrogen-containing heteroaryl ring; or any combinationthereof.
 2. The compound of claim 1, the compound having a formula


3. The compound of claim 1, wherein R^(4a) is deuterium.
 4. The compoundof claim 2, wherein R^(3d) or R^(5a) or both are halogen.
 5. Thecompound of claim 4, wherein R^(3d) or R^(5a) or both are F.
 6. Thecompound of claim 1, wherein the polycyclic is selected from

or adamantly.
 7. The compound of claim 6, wherein the polycyclic is


8. The compound of claim 1, wherein R^(S)C comprises anitrogen-containing heteroaryl ring.
 9. The compound of claim 8, whereinR^(S)C is selected from pyridine, pyrazole, pyrrole, imidazole, oxazole,isoxazole, thiazole, isothiazole, triazole, pyrimidine, pyrazine,triazine, benzopyrazole, benzimidazole, indole, quinoline, indazole,purine, quinoxaline, or acridine.
 10. The compound of claim 1, havingthe formula

wherein Z is selected from N, CH, or C-alkyl; R^(6a), R^(6c), R^(6d) andR^(6g) are each independently selected from H, D, halogen or alkyl; andR^(6h) is selected from H, D, alkyl, cycloalkyl, aryl or heteroaryl. 11.The compound of claim 10, wherein Z is N; R^(6a), R^(6c), R^(6d) andR^(6g) are all H; R^(6h) is methyl; or a combination thereof.
 12. Thecompound of claim 1, having a formula

wherein R^(6a), R^(6b), R^(6c) and R^(6d) are each independentlyselected from H, D, halogen or alkyl; G is a lone pair of electrons, oran oxygen; R^(6e) and R^(6f) are each independently selected from alkyl,H or cycloalkyl; and wherein R^(ad) or R^(y)a or both are halogen; orR^(4a) is D; or R¹ is polycyclic; or any combination thereof.
 13. Thecompound of claim 12, wherein R^(6e) and R^(6f) are both methyl.
 14. Thecompound of claim 1, wherein the compound is


15. A compound, having a formula

or a pharmaceutically acceptable salt thereof, wherein R is selectedfrom

R^(a) is selected from aryl, heteroaryl, alkenyl, cycloalkyl,heterocyclic, or polycyclic; R^(b) is selected from hydrogen, alkyl,alkenyl, or cycloalkyl; Y is CR^(g), N or N—O (N-oxide); R^(c), R^(d),R^(e) and R^(g) are each independently selected from hydrogen,deuterium, halide, alkyl, alkenyl, alkoxy, alkylthio, amino, sulfonyl,aminosulfonyl, aminocarbonyl, acyl, hydroxyl or nitro; R^(fa) and R^(fb)are each independently selected from hydrogen, deuterium, halide oralkyl; L^(a) and L^(b) are each independently selected from hydrogen,deuterium, alkyl or cycloalkyl, or together form a pi-bond; L^(c) andL^(d) are each independently selected from hydrogen, deuterium, alkyl orcycloalkyl; W is selected from O or —(C(L^(c))(L^(d)))_(s)-; s is 1, 2,3, 4, 5 or 6; n is 0 or 1; X is heterocyclic or heteroaryl; and whereinif W is CH₂ and L^(c) and L^(d) are both H, then X is not a benzopyran;if R is

L^(c) and L^(d) are both H, and L^(a) and L^(b) are both H or togetherform a pi-bond, then X is not a benzopyran; X is not substituted with—R^(x)-L^(x)-R^(x2), where R^(x) is selected from O, NR^(x3), sulfonylor S; R^(x3) is selected from H, alkyl, alkenyl, alkynyl, cycloalkyl,cycloalkenyl, or aryl; L^(x) is selected from a bond, alkylene,alkenylene, alkynylene, cycloalkyl, cycloalkenyl, heterocyclic, aryl,heteroaryl or CR^(x4)R^(x5); R^(x4) and R^(x5) are each independentlyselected from H, D, halogen, alkyl, alkenyl, alkynyl, cycloalkyl,cycloalkenyl, —C(O)OR^(x6), or C(O)NR^(x6)R^(x7); R^(x6) and R^(x7) areeach independently selected from H, alkyl, alkenyl, alkynyl, cycloalkylor cycloalkenyl; R^(x2) is selected from —C(O)L^(x2)R^(x8) or a carboxylbioisostere; L^(x2) is a bond or NR^(x3); R^(x8) is H, alkyl, alkenyl,alkynyl, cycloalkyl, cycloalkenyl, —OR^(x9), N(R^(x9))₂, —C(O)R^(x9),—S(O)₂R^(x9), —C(O)OR^(x9), —S(O)₂N(R^(x9))₂ or —C(O)N(R^(x9))₂; andeach R^(x9) is independently selected from H, alkyl, alkenyl, alkynyl,cycloalkyl or cycloalkenyl.
 16. A compound, having a formula

or a pharmaceutically acceptable salt thereof, wherein R^(a) is selectedfrom aryl, heteroaryl, alkyl, alkenyl, cycloalkyl, heterocyclic, orpolycyclic; R^(b) is selected from alkyl, alkenyl, or cycloalkyl; Y isCR^(g), N or N—O (N-oxide); R^(c), R^(d), R^(e) and R^(g) are eachindependently selected from hydrogen, deuterium, halide, alkyl, alkenyl,alkoxy, alkylthio, amino, sulfonyl, aminosulfonyl, aminocarbonyl,cycloalkyl, heterocyclic, acyl, hydroxyl or nitro; R^(fa) and R^(fb) areeach independently selected from hydrogen, deuterium, halide or alkyl;R^(h) and R^(j) are each independently selected from hydrogen,deuterium, halide, alkyl, cycloalkyl, heterocycloalkyl, alkenyl, aryl orheteroaryl; R^(k) and R^(m) are independently selected from H, alkyl,aryl, cycloalkyl, heterocycloalkyl, heteroaryl, or together R^(k) andR^(m) form a cycloalkyl or heterocycloalkyl ring; each R^(n) isindependently selected from H, alkyl, or a metal salt; X is aryl, orheteroaryl; R is selected from

L^(a) and L^(b) are each independently selected from hydrogen,deuterium, alkyl or cycloalkyl, or together form a pi-bond; L^(c) andL^(d) are each independently selected from hydrogen, deuterium, alkyl orcycloalkyl; W is selected from O or —(C(L^(c))(L^(d)))_(s)-; s is 1, 2,3, 4, 5 or 6; n is 0 or 1; R^(y) and R^(z) are selected from alkyl,cycloalkyl, heterocyclic alkyl, aryl, or heteroaryl, or R^(y) and R^(z)may together form a cycloheteroalkyl ring; R^(im) is selected from—OCH₂CO₂Na or

and wherein X is not substituted with —R^(x)-L^(x)-R^(x2), where R^(x)is selected from O, NR^(x3), sulfonyl or S; R^(x3) is selected from H,alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, or aryl; L^(x) isselected from a bond, alkylene, alkenylene, alkynylene, cycloalkyl,cycloalkenyl, heterocyclic, aryl, heteroaryl or CR^(x4)R^(x5); R^(x4)and Rx^(x5) are each independently selected from H, D, halogen, alkyl,alkenyl, alkynyl, cycloalkyl, cycloalkenyl, —C(O)OR^(x6), or—C(O)NR^(x6)R^(x7); R^(x6) and R^(x7) are each independently selectedfrom H, alkyl, alkenyl, alkynyl, cycloalkyl or cycloalkenyl; R^(x2) isselected from —C(O)L^(x2)R^(x8) or a carboxyl bioisostere; L^(x2) is abond or NR^(x3); R^(x8) is H, alkyl, alkenyl, alkynyl, cycloalkyl,cycloalkenyl, —OR^(x9), N(R^(x9))₂, —C(O)R^(x9), —S(O)₂R^(x9),—C(O)OR^(x9), —S(O)₂N(R^(x9))₂ or C(O)N(R^(x9))₂; and each R^(x9) isindependently selected from H, alkyl, alkenyl, alkynyl, cycloalkyl orcycloalkenyl.
 17. A compound, selected from methyl(E)-3-(3-(N-(4-(1-methyl-1H-indazol-5-yl)benzyl)cyclohexanecarboxamido)phenyl)acrylate;methyl(E)-3-(3-((1R,2S,4S)—N-(4-(1-methyl-1H-indazol-5-yl)benzyl)bicyclo[2.2.1]heptane-2-carboxamido)phenyl)acrylate;methyl(E)-3-(3-(1-methyl-N-(4-(1-methyl-1H-indazol-5-yl)benzyl)piperidine-4-carboxamido)phenyl)acrylate;methyl(E)-3-(5-(N-((1-methyl-1H-benzo[f]indazol-8-yl)methyl)cyclohexanecarboxamido)pyridin-3-yl)acrylate;methyl(E)-3-(3-fluoro-5-((1S,2R,4R)—N-((1-methyl-1H-benzo[1]indazol-8-yl)methyl)bicyclo[2.2.1]heptane-2-carboxamido)phenyl)acrylate;methyl(E)-3-(3-(N-((9-fluoro-1-methyl-1H-benzo[f]indazol-8-yl)methyl)cyclohexanecarboxamido)phenyl)acrylate;methyl(E)-3-(3-((1R,4S)—N-((9-fluoro-1-methyl-1H-benzo[f]indazol-8-yl)methyl)bicyclo[2.2.1]heptane-2-carboxamido)phenyl)acrylate;methyl(E)-3-(3-(N-((9-fluoro-1-methyl-1H-benzo[f]indazol-8-yl)methyl)-1-methylpiperidine-4-carboxamido)phenyl)acrylate;methyl(E)-3-(5-(N-((9-fluoro-1-methyl-1H-benzo[f]indazol-8-yl)methyl)cyclohexanecarboxamido)pyridin-3-yl)acrylate;methyl(E)-3-(3-fluoro-5-(N-((9-fluoro-1-methyl-1H-benzo[f]indazol-8-yl)methyl)cyclohexanecarboxamido)phenyl)acrylate;methyl(E)-3-(3-fluoro-5-((1R,4S)—N-((9-fluoro-1-methyl-1H-benzo[f]indazol-8-yl)methyl)bicyclo[2.2.1]heptane-2-carboxamido)phenyl)acrylate;methyl(E)-3-(5-(N-((9-chloro-1-methyl-1H-benzo[f]indazol-8-yl)methyl)cyclohexanecarboxamido)pyridin-3-yl)acrylate;methyl(E)-3-(5-((1R,4S)—N-((9-chloro-1-methyl-1H-benzo[f]indazol-8-yl)methyl)bicyclo[2.2.1]heptane-2-carboxamido)pyridin-3-yl)acrylate;methyl(E)-3-(3-(N-((9-chloro-1-methyl-1H-benzo[f]indazol-8-yl)methyl)-1-methylpiperidine-4-carboxamido)-5-fluorophenyl)acrylate;methyl(E)-3-(3-(N-((7-(dimethylamino)naphthalen-1-yl)methyl)cyclohexanecarboxamido)phenyl)acrylate;methyl(E)-3-(3-(N-((7-(dimethylamino)-8-fluoronaphthalen-1-yl)methyl)cyclohexanecarboxamido)phenyl)acrylate;methyl(E)-3-(3-(N-((4-(1-methyl-1H-indazol-5-yl)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate;methyl(E)-3-(5-(N-((4-(1-methyl-1H-indazol-5-yl)phenyl)methyl-d)cyclohexanecarboxamido)pyridin-3-yl)acrylate;methyl(E)-3-(5-(N-((2-fluoro-4-(1-methyl-1H-indazol-5-yl)phenyl)methyl-d)cyclohexanecarboxamido)pyridin-3-yl)acrylate;methyl(E)-3-(3-fluoro-5-(N-((2-fluoro-4-(1-methyl-1H-indazol-5-yl)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate;methyl(E)-3-(3-(N-((2-chloro-4-(1-methyl-1H-indazol-5-yl)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate;methyl(E)-3-(3-((1S,2R,4R)—N-((2-chloro-4-(1-methyl-1H-indazol-5-yl)phenyl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)phenyl)acrylate;methyl(E)-3-(3-(N-((2-chloro-4-(1-methyl-1H-indazol-5-yl)phenyl)methyl-d)cyclohexanecarboxamido)-5-fluorophenyl)acrylate;methyl(E)-3-(5-((1S,2R,4R)—N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)pyridin-3-yl)acrylate;methyl(E)-3-(3-(N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)cyclohexanecarboxamido)-5-fluorophenyl)acrylate;methyl(E)-3-(3-((1S,2R,4R)—N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)-5-fluorophenyl)acrylate;methyl(E)-3-(3-(N-((4′-(dimethylamino)-3-fluoro-[1,1′-biphenyl]-4-yl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate;methyl(E)-3-(5-(N-((4′-(dimethylamino)-3-fluoro-[1,1′-biphenyl]-4-yl)methyl-d)cyclohexanecarboxamido)pyridin-3-yl)acrylate;methyl(E)-3-(5-((1S,2R,4R)—N-((4′-(dimethylamino)-3-fluoro-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)pyridin-3-yl)acrylate;methyl(E)-3-(3-(N-((3-chloro-4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate;methyl(E)-3-(5-(N-((3-chloro-4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)cyclohexanecarboxamido)pyridin-3-yl)acrylate;methyl(E)-3-(3-fluoro-5-(N-((2-fluoro-4-(1-methyl-1H-indazol-5-yl)phenyl)methyl-d)benzamido)phenyl)acrylate;methyl(E)-3-(3-(N-((2-chloro-4-(1-methyl-1H-indazol-5-yl)phenyl)methyl-d)benzamido)-5-fluorophenyl)acrylate;methyl(E)-3-(3-(N-((2-fluoro-4-(1-methyl-1H-indazol-5-yl)phenyl)methyl-d)benzamido)phenyl)acrylate;methyl(E)-3-(5-((1S,2R,4R)—N-((2-fluoro-4-(1-methyl-1H-indazol-5-yl)phenyl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)pyridin-3-yl)acrylate;methyl(E)-3-(3-(N-(2-chloro-4-(1-methyl-1H-indazol-5-yl)benzyl)-1-methylpiperidine-4-carboxamido)phenyl)acrylate;methyl(E)-3-(3-((1S,2R,4R)—N-(2-chloro-4-(1-methyl-1H-indazol-5-yl)benzyl)bicyclo[2.2.1]heptane-2-carboxamido)phenyl)acrylate;methyl(E)-3-(3-((1S,2R,4R)—N-(2-chloro-4-(1-methyl-1H-indazol-5-yl)benzyl)bicyclo[2.2.1]heptane-2-carboxamido)-5-fluorophenyl)acrylate;methyl(E)-3-(5-((1S,2R,4R)—N-((2-chloro-4-(1-methyl-1H-indazol-5-yl)phenyl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)pyridin-3-yl)acrylate;methyl(E)-3-(3-(N-(4-(1-methyl-1H-indazol-5-yl)benzyl)benzamido)phenyl)acrylate,methyl(E)-3-(3-fluoro-5-(N-(4-(1-methyl-1H-indazol-5-yl)benzyl)benzamido)phenyl)acrylate;methyl(E)-3-(3-(N-(2-fluoro-4-(1-methyl-1H-indazol-5-yl)benzyl)benzamido)phenyl)acrylate;methyl(E)-3-(5-(N-(2-fluoro-4-(1-methyl-1H-indazol-5-yl)benzyl)benzamido)pyridin-3-yl)acrylate;methyl(E)-3-(3-(N-((4-(1-methyl-1H-indazol-5-yl)phenyl)methyl-d)benzamido)phenyl)acrylate;methyl(E)-3-(3-fluoro-5-(N-((4-(1-methyl-1H-indazol-5-yl)phenyl)methyl-d)benzamido)phenyl)acrylate;methyl(E)-3-(5-(N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)cyclohexanecarboxamido)pyridin-3-yl)acrylate;methyl(E)-3-(3-((1S,2R,4R)—N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl)bicyclo[2.2.1]heptane-2-carboxamido)phenyl)acrylate;methyl(E)-3-(5-((1S,2R,4R)—N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl)bicyclo[2.2.1]heptane-2-carboxamido)pyridin-3-yl)acrylate;methyl(E)-3-(3-(N-((3-chloro-4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)cyclohexanecarboxamido)-5-fluorophenyl)acrylate;methyl(E)-3-(3-((1S,2R,4R)—N-((4′-(dimethylamino)-3-fluoro-[1,1′-biphenyl]-4-yl)methyl)bicyclo[2.2.1]heptane-2-carboxamido)phenyl)acrylate;methyl(E)-3-(3-((1S,2R,4R)—N-((4′-(dimethylamino)-3-fluoro-[1,1′-biphenyl]-4-yl)methyl)bicyclo[2.2.1]heptane-2-carboxamido)-5-fluorophenyl)acrylate;methyl(E)-3-(3-((1S,2R,4R)—N-((3-chloro-4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl)bicyclo[2.2.1]heptane-2-carboxamido)phenyl)acrylate;methyl(E)-3-(3-((1S,2R,4R)—N-((3-chloro-4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)phenyl)acrylate;methyl(E)-3-(5-((1S,2R,4R)—N-((3-chloro-4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)pyridin-3-yl)acrylate;methyl(E)-3-(5-(N((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl)benzamido)pyridin-3-yl)acrylate;methyl(E)-3-(3-(N-((4′-(dimethylamino)-3-fluoro-[1,1′-biphenyl]-4-yl)methyl)benzamido)phenyl)acrylate;methyl(E)-3-(3-(N-((3-chloro-4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl)benzamido)phenyl)acrylate;methyl(E)-3-(3-(N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)benzamido)phenyl)acrylate;methyl(E)-3-(5-(N((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)benzamido)pyridin-3-yl)acrylate;methyl(E)-3-(3-(N-((3-chloro-4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)benzamido)-5-fluorophenyl)acrylate;methyl(E)-3-(3-fluoro-5-(N-((4-(1-methyl-1H-indazol-5-yl)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate;methyl(E)-3-(3-fluoro-5-(N-(4-(1-methyl-1H-indazol-5-yl)benzyl)cyclohexanecarboxamido)phenyl)acrylate;methyl(E)-3-(3-(N-((4′-(dimethylamino)-[1,1′biphenyl]-4-yl)methyl)benzamido)-5-fluorophenyl)acrylate;methyl(E)-3-(3-(N-(2-chloro-4-(1-methyl-1H-indazol-5-yl)benzyl)cyclohexanecarboxamido)phenyl)acrylate;methyl(E)-3-(5-((1S,2R,4R)—N-(2-fluoro-4-(1-methyl-1H-indazol-5-yl)benzyl)bicyclo[2.2.1]heptane-2-carboxamido)pyridin-3-yl)acrylate;methyl(E)-3-(3-(N-((2-fluoro-4-(1-methyl-1H-indazol-5-yl)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate;methyl(E)-3-(5-(N-((3-chloro-4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)benzamido)pyridin-3-yl)acrylate;methyl(E)-3-(3-(N-(2-chloro-4-(1-methyl-1H-indazol-5-yl)benzyl)cyclohexanecarboxamido)-5-fluorophenyl)acrylate;methyl(E)-3-(5-(N-((2-chloro-4-(1-methyl-1H-indazol-5-yl)phenyl)methyl-d)cyclohexanecarboxamido)pyridin-3-yl)acrylate;methyl(E)-3-(3-((1S,2R,4R)—N-((2-chloro-4-(1-methyl-1H-indazol-5-yl)phenyl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)-5-fluorophenyl)acrylate;methyl(E)-3-(3-(N-(2-chloro-4-(1-methyl-1H-indazol-5-yl)benzyl)benzamido)-5-fluorophenyl)acrylate;methyl(E)-3-(5-(N-(2-chloro-4-(1-methyl-1H-indazol-5-yl)benzyl)benzamido)pyridin-3-yl)acrylate;methyl(E)-3-(3-fluoro-5-(N-(2-fluoro-4-(1-methyl-1H-indazol-5-yl)benzyl)benzamido)phenyl)acrylate;methyl(E)-3-(3-(N-((2-chloro-4-(1-methyl-1H-indazol-5-yl)phenyl)methyl-d)benzamido)phenyl)acrylate;methyl(E)-3-(5-((1S,2R,4R)—N-((4-(1-methyl-1H-indazol-5-yl)phenyl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)pyridin-3-yl)acrylate;methyl(E)-3-(3-(N-(2-chloro-4-(1-methyl-1H-indazol-5-yl)benzyl)benzamido)phenyl)acrylate;methyl(E)-3-(3-(N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate;methyl(E)-3-(5-(N-((2-chloro-4-(1-methyl-1H-indazol-5-yl)phenyl)methyl-d)benzamido)pyridin-3-yl)acrylate;methyl(E)-3-(3-(N-((3-chloro-4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl)cyclohexanecarboxamido)phenyl)acrylate;methyl(E)-3-(3-(N-((4′-(dimethylamino)-3-fluoro-[1,1′-biphenyl]-4-yl)methyl-d)benzamido)phenyl)acrylate;methyl(E)-3-(5-(N-((4′-(dimethylamino)-3-fluoro-[1,1′-biphenyl]-4-yl)methyl)cyclohexanecarboxamido)pyridin-3-yl)acrylate;methyl(E)-3-(5-((1S,2R,4R)—N-((3-chloro-4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl)bicyclo[2.2.1]heptane-2-carboxamido)pyridin-3-yl)acrylate;methyl(E)-3-(3-fluoro-5-((1S,2R,4R)—N-((4-(1-methyl-1H-indazol-5-yl)phenyl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)phenyl)acrylate;methyl(E)-3-(3-((1S,2R,4R)—N-((4-(1-methyl-1H-indazol-5-yl)phenyl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)phenyl)acrylate;methyl(E)-3-(5-(N-((3-chloro-4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl)benzamido)pyridin-3-yl)acrylate;methyl(E)-3-(3-((1S,2R,4R)—N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl)bicyclo[2.2.1]heptane-2-carboxamido)-5-fluorophenyl)acrylate;methyl(E)-3-(3-((1S,2R,4R)—N-((2-fluoro-4-(1-methyl-1H-indazol-5-yl)phenyl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)phenyl)acrylate;methyl(E)-3-(3-(N-((4′-(dimethylamino)-3-fluoro-[1,1′-biphenyl]-4-yl)methyl-d)cyclohexanecarboxamido)-5-fluorophenyl)acrylate;methyl(E)-3-(3-(N-((3-chloro-4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl)benzamido)-5-fluorophenyl)acrylate;methyl(E)-3-(3-((1S,2R,4R)—N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)phenyl)acrylate;methyl(E)-3-(3-((1S,2R,4R)—N-((4′-(dimethylamino)-3-fluoro-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)-5-fluorophenyl)acrylate;methyl(E)-3-(3-(N-((4′-(dimethylamino)-3-fluoro-[1,1′-biphenyl]-4-yl)methyl-d)benzamido)-5-fluorophenyl)acrylate;methyl(E)-3-(3-(N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)benzamido)-5-fluorophenyl)acrylate;methyl(E)-3-(3-((1S,2R,4R)—N-((3-chloro-4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)-5-fluorophenyl)acrylate;methyl(E)-3-(5-(N-(4-(1-methyl-1H-indazol-5-yl)benzyl)benzamido)pyridin-3-yl)acrylate;methyl(E)-3-(5-(N-((4-(1-methyl-1H-indazol-5-yl)phenyl)methyl-d)benzamido)pyridin-3-yl)acrylate;methyl(E)-3-(3-(N-((3-chloro-4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl)cyclohexanecarboxamido)-5-fluorophenyl)acrylate;methyl(E)-3-(5-(N-((4′-(dimethylamino)-3-fluoro-[1,1′-biphenyl]-4-yl)methyl-d)benzamido)pyridin-3-yl)acrylate;methyl(E)-3-(5-((1S,2R,4R)—N-((4′-(dimethylamino)-3-fluoro-[1,1′-biphenyl]-4-yl)methyl)bicyclo[2.2.1]heptane-2-carboxamido)pyridin-3-yl)acrylate;methyl(E)-3-(3-(N-(4-(2-(tert-butoxy)-2-oxoethoxy)benzyl)cyclohexanecarboxamido)phenyl)acrylate;tert-butyl (E)-2-(4-((N-(3-(3-methoxy-3-oxoprop-1-en-1-yl)phenyl)cyclohexanecarboxamido)methyl)phenyl)cyclopropane-1-carboxylate;methyl(E)-3-(3-(N-(4-((2-oxotetrahydro-2H-pyran-3-yl)methyl)benzyl)cyclohexanecarboxamido)phenyl)acrylate;cyclobutyl(E)-3-(4-((N-(3-((E)-3-methoxy-3-oxoprop-1-en-1-yl)phenyl)cyclohexanecarboxamido)methyl)phenyl)acrylate;methyl(E)-3-(3-(N-(4-(6-methoxypyridin-3-yl)benzyl)cyclohexanecarboxamido)phenyl)acrylate;methyl(E)-3-(3-(N-((5-(4-(dimethylamino)phenyl)pyridin-2-yl)methyl)cyclohexanecarboxamido)phenyl)acrylate;methyl(E)-3-(3-(N-((4-(4-(dimethylamino)phenyl)piperazin-1-yl)methyl)cyclohexanecarboxamido)phenyl)acrylate;methyl(E)-3-(3-(N-(4-(3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)benzyl)cyclohexanecarboxamido)phenyl)acrylate;methyl(E)-3-(3-(N-(4-(2H-benzo[b][1,4]oxazin-7-yl)benzyl)cyclohexanecarboxamido)phenyl)acrylate;methyl(E)-3-(3-(N-(4-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)benzyl)cyclohexanecarboxamido)phenyl)acrylate;methyl(E)-3-(3-(N-(4-(quinoxalin-6-yl)benzyl)cyclohexanecarboxamido)phenyl)acrylatemethyl(E)-3-(3-(N-(4-(1,2,3,4-tetrahydroquinoxalin-6-yl)benzyl)cyclohexanecarboxamido)phenyl)acrylate;methyl(E)-3-(3-(N-(4-(3,4-dihydro-2H-benzo[b][1,4]thiazin-7-yl)benzyl)cyclohexanecarboxamido)phenyl)acrylate;methyl(E)-3-(3-(N-(4-(1,1-dioxido-3,4-dihydro-2H-benzo[b][1,4]thiazin-7-yl)benzyl)cyclohexanecarboxamido)phenyl)acrylate;methyl(E)-3-(3-(N-(4-(1,1,4,4-tetraoxido-2,3-dihydrobenzo[b][1,4]dithiin-6-yl)benzyl)cyclohexanecarboxamido)phenyl)acrylate;methyl2-(3-(N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl)cyclohexanecarboxamido)phenyl)cyclopropane-1-carboxylate;methyl2-(3-(N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl)cyclohexanecarboxamido)phenyl)cyclobutane-1-carboxylate;methyl2-(3-(N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl)cyclohexanecarboxamido)phenyl)cyclopentane-1-carboxylate;methyl2-(3-(N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl)cyclohexanecarboxamido)phenyl)cyclohexane-1-carboxylate;methyl(E)-3-(3-(N-(4-(benzo[d]oxazol-6-yl)benzyl)cyclohexanecarboxamido)phenyl)acrylate;methyl(E)-3-(3-(N-(4-(1H-benzo[d]imidazol-6-yl)benzyl)cyclohexanecarboxamido)phenyl)acrylate;methyl(E)-3-(3-(N-(4-(benzo[d]thiazol-6-yl)benzyl)cyclohexanecarboxamido)phenyl)acrylate;methyl(E)-3-(3-(N-(4-(2-methylbenzo[d]thiazol-6-yl)benzyl)cyclohexanecarboxamido)phenyl)acrylate;methyl(E)-3-(3-(N-(4-(2-methylbenzo[d]oxazol-6-yl)benzyl)cyclohexanecarboxamido)phenyl)acrylate;methyl(E)-3-(3-(N-(4-(2-methyl-1H-benzo[d]imidazol-6-yl)benzyl)cyclohexanecarboxamido)phenyl)acrylate;methyl(E)-3-(3-(N-(4-(1,2-dimethyl-1H-benzo[d]imidazol-6-yl)benzyl)cyclohexanecarboxamido)phenyl)acrylate;methyl(E)-3-(3-(N-(4-(1-methyl-1H-benzo[d]imidazol-6-yl)benzyl)cyclohexanecarboxamido)phenyl)acrylate;methyl(E)-3-(3-(N-(4-(benzo[d]isoxazol-5-yl)benzyl)cyclohexanecarboxamido)phenyl)acrylate;methyl(E)-3-(3-(N-(4-(benzo[d]isothiazol-5-yl)benzyl)cyclohexanecarboxamido)phenyl)acrylate;methyl3-(3-(N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl)bicyclo[2.2.1]heptane-2-carboxamido)phenyl)propanoate;methyl2-(3-(N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl)bicyclo[2.2.1]heptane-2-carboxamido)phenoxy)acetate;3-(N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl)bicyclo[2.2.1]heptane-2-carboxamido)benzylacetate;N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl)-N-(3-((2-oxotetrahydro-2H-pyran-3-yl)methyl)phenyl)cyclohexanecarboxamide;N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl)-N-(3-((2-oxotetrahydro-2H-pyran-4-yl)methyl)phenyl)cyclohexanecarboxamide;methyl(E)-3-(3-(N-((4-(2-(tert-butoxy)-2-oxoethoxy)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate;tert-butyl(E)-2-(4-((N-(3-(3-methoxy-3-oxoprop-1-en-1-yl)phenyl)cyclohexanecarboxamido)methyl-d)phenyl)cyclopropane-1-carboxylate;methyl(E)-3-(3-(N-((4-((2-oxotetrahydro-2H-pyran-3-yl)methyl)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate;cyclobutyl(E)-3-(4-((N-(3-((E)-3-methoxy-3-oxoprop-1-en-1-yl)phenyl)cyclohexanecarboxamido)methyl-d)phenyl)acrylate;methyl(E)-3-(3-(N-((4-(6-methoxypyridin-3-yl)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate;methyl(E)-3-(3-(N-((5-(4-(dimethylamino)phenyl)pyridin-2-yl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate;methyl(E)-3-(3-(N-((4-(4-(dimethylamino)phenyl)piperazin-1-yl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate;methyl(E)-3-(3-(N-((4-(3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate;methyl(E)-3-(3-(N-((4-(2H-benzo[b][1,4]oxazin-7-yl)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate;methyl(E)-3-(3-(N-((4-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate;methyl(E)-3-(3-(N-((4-(quinoxalin-6-yl)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate;methyl(E)-3-(3-(N-((4-(1,2,3,4-tetrahydroquinoxalin-6-yl)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate;methyl(E)-3-(3-(N-((4-(3,4-dihydro-2H-benzo[b][1,4]thiazin-7-yl)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate;methyl(E)-3-(3-(N-((4-(1,1-dioxido-3,4-dihydro-2H-benzo[b][1,4]thiazin-7-yl)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate;methyl(E)-3-(3-(N-((4-(1,1,4,4-tetraoxido-2,3-dihydrobenzo[b][1,4]dithiin-6-yl)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate;methyl2-(3-(N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)cyclohexanecarboxamido)phenyl)cyclopropane-1-carboxylate;methyl2-(3-(N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)cyclohexanecarboxamido)phenyl)cyclobutane-1-carboxylate;methyl2-(3-(N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)cyclohexanecarboxamido)phenyl)cyclopentane-1-carboxylate;methyl2-(3-(N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)cyclohexanecarboxamido)phenyl)cyclohexane-1-carboxylate;methyl(E)-3-(3-(N-((4-(benzo[d]oxazol-6-yl)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate;methyl(E)-3-(3-(N-((4-(1H-benzo[d]imidazol-6-yl)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate;methyl(E)-3-(3-(N-((4-(benzo[d]thiazol-6-yl)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate;methyl(E)-3-(3-(N-((4-(2-methylbenzo[d]thiazol-6-yl)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate;methyl(E)-3-(3-(N-((4-(2-methylbenzo[d]oxazol-6-yl)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate;methyl(E)-3-(3-(N-((4-(2-methyl-1H-benzo[d]imidazol-6-yl)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate;methyl(E)-3-(3-(N-((4-(1,2-dimethyl-1H-benzo[d]imidazol-6-yl)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate;methyl(E)-3-(3-(N-((4-(1-methyl-1H-benzo[d]imidazol-6-yl)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate;methyl(E)-3-(3-(N-((4-(benzo[d]isoxazol-5-yl)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate;methyl(E)-3-(3-(N-((4-(benzo[d]isothiazol-5-yl)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate;methyl3-(3-(N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)phenyl)propanoate;methyl2-(3-(N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)phenoxy)acetate;3-(N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)benzylacetate;N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)-N-(3-((2-oxotetrahydro-2H-pyran-3-yl)methyl)phenyl)cyclohexanecarboxamide;N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)-N-(3-((2-oxotetrahydro-2H-pyran-4-yl)methyl)phenyl)cyclohexanecarboxamide;methyl(E)-3-(3-((1S,2R,4R)—N—((S)-(4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)-5-fluorophenyl)acrylate;methyl(E)-3-(3-((1S,2R,4R)—N—((R)-(4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)-5-fluorophenyl)acrylate;methyl(E)-3-(3-((1S,2S,4R)—N—((S)-(4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)-5-fluorophenyl)acrylate;methyl(E)-3-(3-((1S,2S,4R)—N—((R)-(4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)-5-fluorophenyl)acrylate;methyl(E)-3-(3-((1R,2S,4S)—N—(S)-(4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)-5-fluorophenyl)acrylate;methyl(E)-3-(3-((1R,2S,4S)—N—((R)-(4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)-5-fluorophenyl)acrylate;methyl(E)-3-(3-((1R,2R,4S)—N—((S)-(4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)-5-fluorophenyl)acrylate;methyl(E)-3-(3-((1R,2R,4S)—N—((R)-(4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)-5-fluorophenyl)acrylate;methyl(E)-3-(3-((1S,2R,4R)—N—((S)-(4′-(dimethylamino)-3-fluoro-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)-5-fluorophenyl)acrylate;methyl(E)-3-(3-((1S,2R,4R)—N—((R)-(4′-(dimethylamino)-3-fluoro-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)-5-fluorophenyl)acrylate;methyl(E)-3-(3-((1S,2S,4R)—N—((S)-(4′-(dimethylamino)-3-fluoro-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)-5-fluorophenyl)acrylate;methyl(E)-3-(3-((1S,2S,4R)—N—((R)-(4′-(dimethylamino)-3-fluoro-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)-5-fluorophenyl)acrylate;methyl(E)-3-(3-((1R,2S,4S)—N—((S)-(4′-(dimethylamino)-3-fluoro-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)-5-fluorophenyl)acrylate,methyl(E)-3-(3-((1R,2S,4S)—N—((R)-(4′-(dimethylamino)-3-fluoro-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)-5-fluorophenyl)acrylate;methyl(E)-3-(3-((1R,2R,4S)—N—((S)-(4′-(dimethylamino)-3-fluoro-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)-5-fluorophenyl)acrylate;methyl(E)-3-(3-((1R,2R,4S)—N—((R)-(4′-(dimethylamino)-3-fluoro-[1,1′-biphenyl]-4-yl)methyl-d)bicyclo[2.2.1]heptane-2-carboxamido)-5-fluorophenyl)acrylate;methyl(R,E)-3-(3-(N-((4′-(dimethylamino)-3-fluoro-[1,1′-biphenyl]-4-yl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate;methyl(S,E)-3-(3-(N-((4′-(dimethylamino)-3-fluoro-[1,1′-biphenyl]-4-yl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate;methyl(R,E)-3-(3-(N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate,methyl(S,E)-3-(3-(N-((4′-(dimethylamino)-[1,1′-biphenyl]-4-yl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate;methyl(R,E)-3-(3-fluoro-5-(N-((4-(1-methyl-1H-indazol-5-yl)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate;methyl(S,E)-3-(3-fluoro-5-(N-((4-(1-methyl-1H-indazol-5-yl)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate;methyl(R,E)-3-(3-(N-((2-fluoro-4-(1-methyl-1H-indazol-5-yl)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate;or methyl(S,E)-3-(3-(N-((2-fluoro-4-(1-methyl-1H-indazol-5-yl)phenyl)methyl-d)cyclohexanecarboxamido)phenyl)acrylate.18. A method of making a compound of claim 1, the method comprising:reacting an aldehyde with a first amine to form an imine; reacting theimine with a reducing agent to form a second amine; and reacting thesecond amine with an activated carboxylic acid derivative or acarboxylic acid to form an amide.
 19. A composition, comprising acompound of claim 1 and an additional component.
 20. A method oftreating or preventing a disorder or disease in a subject, comprisingadministering to a gastrointestinal tract of the subject atherapeutically effective amount of a compound of claim
 1. 21. Themethod of claim 20, wherein the disorder or disease is selected from ametabolic disorder, inflammation in an intestinal region, a cellproliferation disease, alcoholic liver disease, non-alcoholic liverdisease, a cholestatic disorder, an intestinal permeability disorder, adisease that causes or results from an altered intestinal microbiome, aninborn error of metabolism, a bile disorder, or an intestinalmalabsorption disorder.
 22. The method of claim 20, wherein the disorderor disease is selected from obesity, diabetes, insulin resistance,dyslipidemia, necrotizing enterocolitis, gastritis, ulcerative colitis,Crohn's disease, inflammatory bowel disease, irritable bowel syndrome,gastroenteritis, radiation induced enteritis, pseudomembranous colitis,chemotherapy induced enteritis, gastro-esophageal reflux disease (GERD),peptic ulcer, non-ulcer dyspepsia (NUD), celiac disease, intestinalceliac disease, colon cancer, post-surgical inflammation, gastriccarcinogenesis, fatty liver (steatosis), cirrhosis, alcoholic hepatitis,nonalcoholic steatohepatitis (NASH), or nonalcoholic fatty liver disease(NAFLD), primary biliary cirrhosis (PBC), primary sclerosing cholangitis(PSC), cholestasis resulting from a drug, overlap syndrome (PBC plusautoimmune hepatitis), drug-induced cholestatic hepatitis, totalparenteral nutrition (TPN)-induced cholestasis, ICU/sepsis-relatedcholestasis, obstetric cholestasis, graft vs. host disease, prolongedcholestasis due to hepatitis A, B or C infection, cholestasis due tocystic fibrosis, alcoholic hepatitis, progressive familial intrahepaticcholestasis (PFIC) syndromes, Alagille syndrome, biliary atresia,infectious colitis, type 1 diabetes, an intestinal permeabilitycondition, an intestinal inflammation disorder, intestinal lesions,cirrhosis, cerebrotendinous xanthomatosis (CTX), benign biliarystricture, malignant biliary obstruction, bile acid diarrhea, shortbowel syndrome, tropical sprue, environmental enteropathy, or volvulus.23. A method of treating or preventing a disorder or disease in asubject, comprising administering to a gastrointestinal tract of thesubject a therapeutically effective amount of a composition of claim 19.24. The method of claim 23, wherein the composition comprises an entericcoating.